Fungal textile materials and leather analogs

ABSTRACT

Textile compositions comprising at least one filamentous fungus are disclosed, as are methods for making and using such textile compositions. Embodiments of the textile compositions generally include at least one of a plasticizer, a polymer, and a crosslinker, in addition to the filamentous fungus. The disclosed textile compositions are particularly useful as analogs or substitutes for conventional textile compositions, including but not limited to leather.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.17/030,322, filed 23 Sep. 2020, which is a continuation of U.S. patentapplication Ser. No. 16/904,520, filed 17 Jun. 2020, which claims thebenefit of priority of U.S. Provisional Patent Applications 62/862,680,filed 18 Jun. 2019; 62/951,332, filed 20 Dec. 2019; and 62/966,525,filed 27 Jan. 2020. All of the above-referenced applications areincorporated by reference herein in their entireties.

FIELD OF THE INVENTION

This invention relates generally to fungal materials, and particularlyto materials derived from filamentous fungi that can be used as leatheranalogs and in other textiles and fabrics.

BACKGROUND OF THE INVENTION

Many current textile materials, including but not limited to leather,create environmental problems during manufacturing and may be difficultor impossible to recycle or dispose of in an environmentally safe way atthe end of an article's useful life. By way of non-limiting example, themanufacture of leather depends on the rearing of cattle (which has asignificant environmental impact in itself and may raise animal welfareconcerns) and requires a tanning step, which may use highly toxicchemicals such as chromium, formic acid, mercury, and various solvents.Leather also biodegrades slowly, over times of about 25 to about 40years. Many textile materials suffer from similar environmental orethical concerns.

There is thus a need in the art for textile materials that may beproduced cost-effectively with a minimum of environmental impact andwithout animal welfare or other ethical concerns. It is furtheradvantageous for such materials to retain various physical and/ormechanical properties, e.g. tensile strength, tear strength, flexuralrigidity, elasticity, texture, thermal properties, sensory attributes,etc., of conventional textile materials, e.g. leather.

SUMMARY OF THE INVENTION

In one aspect of the present invention, a method for preparing a durablesheet material comprising fungal biomass comprises (a) causing asolution to infiltrate an inactivated fungal biomass, the solutioncomprising a solvent and a component selected from the group consistingof a polymer, a crosslinker, and combinations and mixtures thereof; and(b) curing the biomass to remove solvent from the biomass and form thedurable sheet material.

In embodiments, the fungal biomass may comprise fungal mycelia.

In embodiments, the inactivated fungal biomass may be size-reduced priorto step (a) and step (a) may comprise blending the size-reducedinactivated fungal biomass with the solution to form a blendedcomposition. The method may, but need not, further comprise, casting theblended composition to form a cast sheet from which solvent is removedin step (b). The size-reduced inactivated fungal biomass may, but neednot, have an average particle size of no more than about 125 microns.

In embodiments, the inactivated fungal biomass may comprise a cohesivefungal biomass and (a) may comprise agitating the inactivated fungalbiomass and the solution together for a time period. The cohesive fungalbiomass may, but need not, be produced by a surface fermentation processor a submerged solid surface fermentation process. The time period may,but need not, be selected from the group consisting of at least about 4hours, at least about 5 hours, at least about 10 hours, at least about15 hours, at least about 20 hours, or at least about 25 hours. The timeperiod may, but need not, be between about 10 hours and about 20 hours.The agitating may, but need not, be carried out at a pressure other thanatmospheric pressure, which may be sub-atmospheric pressure orsuper-atmospheric pressure. The method may, but need not, furthercomprise subjecting the inactivated fungal biomass to treatment with atleast one chemical selected from the group consisting of calciumhydroxide and tannins.

In embodiments, the inactivated fungal biomass may comprise a fungalpaste produced by submerged fermentation.

In embodiments, the polymer may be selected from the group consisting ofpolyvinyl alcohol, chitosan, polyethylene glycol, alginates, starches,polycaprolactones, polyacrylic acids, hyaluronic acid, and combinationsthereof.

In embodiments, the polymer may be present in the durable sheet materialin an amount selected from the group consisting of no more than about 25wt % of the durable sheet material, no more than about 20 wt % of thedurable sheet material, no more than about 15 wt % of the durable sheetmaterial, no more than about 10 wt % of the durable sheet material, andno more than about 5 wt % of the durable sheet material.

In embodiments, the crosslinker may be selected from the groupconsisting of citric acid, tannic acid, suberic acid, adipic acid,succinic acid, extracted vegetable tannins, glyoxal, and combinationsthereof.

In embodiments, the solution may further comprise a plasticizer. Theplasticizer may, but need not, be selected from the group consisting ofglycerol and esters thereof, polyethylene glycol, citric acid, oleicacid, oleic acid polyols and esters thereof, epoxidized triglyceridevegetable oils, castor oil, pentaerythritol, fatty acid esters,carboxylic ester-based plasticizers, trimellitates, adipates, sebacates,maleates, biological plasticizers, and combinations thereof.

In embodiments, the fungal biomass may comprise at least one filamentousfungus belonging to an order selected from the group consisting ofUstilaginales, Russulales, Agaricales, Pezizales, and Hypocreales.

In embodiments, the fungal biomass may comprise at least one filamentousfungus belonging to a family selected from the group consisting ofUstilaginaceae, Hericiaceae, Polyporaceae, Grifolaceae, Lyophyllaceae,Strophariaceae, Lycoperdaceae, Agaricaceae, Pleurotaceae,Physalacriaceae, Omphalotaceae, Tuberaceae, Morchellaceae,Sparassidaceae, Nectriaceae, and Cordycipitaceae.

In embodiments, the fungal biomass may comprise at least one filamentousfungus belonging to a genus selected from the group consisting ofAgaricus, Calocybe, Calvatia, Cordyceps, Disciotis, Fomes, Fusarium,Ganoderma, Grifola, Hericulum, Hypholoma, Hypsizygus, Morchella,Pholiota, Pleurotus, Polyporous, Sparassis, Stropharia, Tuber, andUstilago.

In embodiments, the fungal biomass may comprise at least one filamentousfungus selected from the group consisting of Ustilago esculenta,Hericulum erinaceus, Polyporous squamosus, Grifola fondosa, Hypsizygusmarmoreus, Hypsizygus ulmarius, Calocybe gambosa, Pholiota nameko,Calvatia gigantea, Agaricus bisporus, Stropharia rugosoannulata,Hypholoma lateritium, Pleurotus eryngii, Pleurotus ostreatus, Pleurotusostreatus var. columbinus, Tuber borchii, Morchella esculenta, Morchellaconica, Morchella importuna, Sparassis crispa, Fusarium venenatum, MK7ATCC Accession Deposit No. PTA-10698, Disciotis venosa, and Cordycepsmilitaris.

In embodiments, the solution may further comprise at least one of apigment, a solubilizer, and a pH adjusting agent. The solubilizer may,but need not, be selected from the group consisting of hydrochloricacid, acetic acid, formic acid, lactic acid, and combinations andmixtures thereof. The pH adjusting agent may, but need not, be selectedfrom the group consisting of hydrochloric acid, acetic acid, formicacid, lactic acid, and combinations and mixtures thereof.

In embodiments, the durable sheet material may comprise proteinscrosslinked with isopeptide bonds.

In embodiments, the method may further comprise at least one of (i)adding a thermal dopant to the inactivated fungal biomass and (ii)adding a thermal dopant to the durable sheet material after step (b). Anamount of the thermal dopant may, but need not, be selected from thegroup consisting of at least about 2.5 wt % of the durable sheetmaterial, at least about 5 wt % of the durable sheet material, at leastabout 7.5 wt % of the durable sheet material, at least about 10 wt % ofthe durable sheet material, at least about 12.5 wt % of the durablesheet material, at least about 15 wt % of the durable sheet material,and at least about 17.5 wt % of the durable sheet material. An amount ofthe thermal dopant may, but need not, be selected from the groupconsisting of no more than about 20 wt % of the durable sheet material,no more than about 17.5 wt % of the durable sheet material, no more thanabout 15 wt % of the durable sheet material, no more than about 12.5 wt% of the durable sheet material, no more than about 10 wt % of thedurable sheet material, no more than about 7.5 wt % of the durable sheetmaterial, and no more than about 5 wt % of the durable sheet material.The thermal dopant may, but need not, be selected from the groupconsisting of a ceramic material, a metallic material, a polymericmaterial, and combinations thereof. The thermal dopant may, but neednot, be selected from the group consisting of activated charcoal,aluminum oxide, bentonite, diatomaceous earth, ethylene vinyl acetate,lignin, nanosilica, polycaprolactone, polylactic acid, silicone, andyttrium oxide.

In embodiments, the inactivated fungal biomass may be a size-reducedinactivated fungal biomass.

In embodiments, the inactivated fungal biomass may comprise a biomat, orportion thereof, produced by a surface fermentation process. Acarbon-to-nitrogen molar ratio in a growth medium of the surfacefermentation process may, but need not, be between about 5 and about 20,or between about 7 and about 15.

In another aspect of the present invention, a textile compositioncomprises an inactivated fungal biomass; and at least one componentselected from the group consisting of a plasticizer, a polymer, acrosslinker, and a dye, wherein the at least one component isdistributed in the fungal mycelial biomass.

In embodiments, the textile composition may have a thickness of at leastabout 1 mm.

In embodiments, the textile composition may have a tear force of atleast about 30 N.

In embodiments, the textile composition may have a tear strength of atleast about 10 N/mm.

In embodiments, the textile composition may have a flexural rigidity ofno more than about 5 gram-centimeters.

In embodiments, the textile composition may have a tensile strength ofat least about 10 MPa.

In embodiments, the textile composition may have a water spotting greyscale rating of at least about 3.

In embodiments, the textile composition may have a light color fastnessblue wool rating of at least about 4.

In embodiments, the textile composition may have a rub color fastnessgrey scale rating, when dry, of at least about 3.

In embodiments, the textile composition may have a rub color fastnessgrey scale rating, when wet, of at least about 2.

In embodiments, the textile composition may further comprise at leastone backing layer of a non-fungal textile material. The non-fungaltextile material may, but need not, be selected from the groupconsisting of an acrylic textile, an alpaca textile, an angora textile,a cashmere textile, a coir textile, a cotton textile, an eisengarntextile, a hemp textile, a jute textile, a Kevlar textile, a linentextile, a microfiber textile, a mohair textile, a nylon textile, anolefin textile, a pashmina textile, a polyester textile, a piña textile,a ramie textile, a rayon textile, a sea silk textile, a silk textile, asisal textile, a spandex textile, a spider silk textile, a wool textile,and combinations and mixtures thereof.

In embodiments, the textile composition may further comprise a thermaldopant. T thermal dopant may, but need not, be selected from the groupconsisting of a ceramic material, a metallic material, a polymericmaterial, and combinations and mixtures thereof. The thermal dopant may,but need not, be selected from the group consisting of activatedcharcoal, aluminum oxide, bentonite, diatomaceous earth, ethylene vinylacetate, lignin, nanosilica, polycaprolactone, polylactic acid,silicone, and yttrium oxide. A thermal characteristic of the textilecomposition may, but need not, be altered relative to the same thermalcharacteristic of the textile composition in the absence of the thermaldopant, wherein the thermal characteristic is selected from the groupconsisting of thermal effusivity, thermal conductivity, heat capacity,and combinations thereof.

In another aspect of the present invention, an article of manufacturecomprises a textile composition as described herein, wherein the articleof manufacture is selected from the group consisting of an article ofclothing, an accessory item, and a furniture item.

In another aspect of the present invention, a method for making adurable sheet material comprises (a) contacting an inactivated fungalbiomass with an aqueous solution comprising calcium hydroxide to form alimed inactivated fungal biomass; (b) contacting the limed inactivatedfungal biomass with an aqueous solution comprising ammonium sulfate toform a delimed inactivated fungal biomass; (c) contacting the delimedinactivated fungal biomass with an aqueous solution comprising a polymerto form a pickled inactivated fungal biomass; (d) contacting the pickledinactivated fungal biomass with an aqueous solution comprising acrosslinker to form a tanned inactivated fungal biomass; (e) contactingthe tanned inactivated fungal biomass with an aqueous solutioncomprising a plasticizer to form a plasticized inactivated fungalbiomass; (f) drying the plasticized inactivated fungal biomass to form adried inactivated fungal biomass; and (g) heat-pressing the driedinactivated fungal biomass to form the durable sheet material.

In embodiments, the method may further comprise, between any pair ofsteps selected from the group consisting of steps (a) and (b), steps (b)and (c), steps (c) and (d), and steps (d) and (e), rinsing theinactivated fungal biomass with water to remove residual aqueoussolution.

In embodiments, at least one of steps (a) through (e) may compriseagitating the inactivated fungal biomass with the aqueous solution.

In embodiments, the aqueous solution of at least one of steps (a)through (c) may further comprise a surfactant or solubilizer. Thesurfactant or solubilizer may, but need not, be selected from the groupconsisting of poly sorbates, hydrochloric acid, acetic acid, formicacid, lactic acid, and combinations and mixtures thereof.

In embodiments, the polymer may be selected from the group consisting ofpolyvinyl alcohol, chitosan, polyethylene glycol, alginates, starches,polycaprolactones, polyacrylic acids, hyaluronic acid, and combinationsand mixtures thereof.

In embodiments, the aqueous solution of step (c) may further comprise aplasticizer selected from the group consisting of glycerol and estersthereof, polyethylene glycol, citric acid, oleic acid, oleic acidpolyols and esters thereof, epoxidized triglyceride vegetable oils,castor oil, pentaerythritol, fatty acid esters, carboxylic ester-basedplasticizers, trimellitates, adipates, sebacates, maleates, biologicalplasticizers, and combinations and mixtures thereof.

In embodiments, the aqueous solution of step (c) may further comprise analkali metal halide. The alkali metal halide may, but need not, besodium chloride.

In embodiments, the crosslinker may be selected from the groupconsisting of citric acid, tannic acid, suberic acid, adipic acid,succinic acid, extracted vegetable tannins, glyoxal, and combinationsand mixtures thereof.

In embodiments, the plasticizer may be selected from the groupconsisting of glycerol and esters thereof, polyethylene glycol, citricacid, oleic acid, oleic acid polyols and esters thereof, epoxidizedtriglyceride vegetable oils, castor oil, pentaerythritol, fatty acidesters, carboxylic ester-based plasticizers, trimellitates, adipates,sebacates, maleates, biological plasticizers, and combinations andmixtures thereof.

In another aspect of the present invention, a method for making adurable sheet material comprises (a) inactivating a fungal biomass byboiling the biomass in water; (b) contacting the inactivated fungalbiomass with an aqueous solution comprising calcium hydroxide to form alimed inactivated fungal biomass; (c) contacting the limed inactivatedfungal biomass with an aqueous solution comprising ammonium sulfate toform a delimed inactivated fungal biomass; (d) contacting the delimedinactivated fungal biomass with an aqueous solution comprising an alkalimetal halide to form a pickled inactivated fungal biomass; (e)contacting the pickled inactivated fungal biomass with a firstcrosslinker to form a tanned inactivated fungal biomass; (f) contactingthe tanned inactivated fungal biomass with an aqueous solutioncomprising at least one of a second crosslinker and a polymer to form are-tanned inactivated fungal biomass; (g) contacting the re-tannedinactivated fungal biomass with a fatliquoring oil to form a fatliquoredinactivated fungal biomass; (h) adhering a non-fungal textile backing tothe inactivated fungal biomass to form a backed inactivated fungalbiomass; (i) heat-pressing the backed inactivated fungal biomass to forma heat-pressed inactivated fungal biomass; (j) drying the heat-pressedinactivated fungal biomass to form a dried inactivated fungal biomass;and (k) applying at least one of a finishing wax, a finishing oil, andnitrocellulose to the dried inactivated fungal biomass to form thedurable sheet material.

In embodiments, the method may further comprise, between any pair ofsteps selected from the group consisting of steps (b) and (c), steps (c)and (d), and steps (e) and (f), rinsing the inactivated fungal biomasswith water to remove residual aqueous solution.

In embodiments, at least one of steps (a) through (g) may compriseagitating the inactivated fungal biomass with the aqueous solution.

In embodiments, the aqueous solution of at least one of steps (b) and(c) may further comprise a surfactant or solubilizer. The surfactant orsolubilizer may, but need not, be selected from the group consisting ofpoly sorbates, hydrochloric acid, acetic acid, formic acid, lactic acid,and combinations and mixtures thereof.

In embodiments, the polymer may, but need not, be selected from thegroup consisting of polyvinyl alcohol, chitosan, polyethylene glycol,alginates, starches, polycaprolactones, polyacrylic acids, hyaluronicacid, and combinations and mixtures thereof

In embodiments, the alkali metal halide may be sodium chloride.

In embodiments, the aqueous solution of at least one of steps (d)through (f) may comprise a pH adjusting agent. The pH adjusting agentmay, but need not, comprise hydrochloric acid, acetic acid, formic acid,lactic acid, or a combination or mixture thereof, or a metal hydroxide.

In embodiments, the first crosslinker may comprise an aluminum salt, achromium salt, a titanium salt, an aldehyde, or a combination or mixturethereof. The first crosslinker may, but need not, be an aluminumsilicate.

In embodiments, the second crosslinker may be selected from the groupconsisting of citric acid, tannic acid, suberic acid, adipic acid,succinic acid, extracted vegetable tannins, glyoxal, and combinationsand mixtures thereof.

In embodiments, the polymer may be selected from the group consisting ofpolyvinyl alcohol, chitosan, polyethylene glycol, alginates, starches,polycaprolactones, polyacrylic acids, hyaluronic acid, and combinationsand mixtures thereof.

In embodiments, the aqueous solution of step (f) may further comprise ananionic dye.

In embodiments, the fatliquoring oil may be selected from the groupconsisting of sulfated castor oil, beeswax, coconut oil, vegetable oil,olive oil, linseed oil, oleic acid, and combinations and mixturesthereof.

In embodiments, the fatliquoring oil may comprise an emulsion and themethod may further comprise, between steps (g) and (h), contacting thefatliquoring oil with an acid to dissociate the emulsion.

In embodiments, the finishing wax may be selected from the groupconsisting of carnauba wax, candelilla wax, and combinations andmixtures thereof.

Embodiments of the present invention generally relate to production ofdurable sheet materials comprising fungal biomass. In certainembodiments, durable sheet materials may have controlled, engineered,and/or tuned thermal properties. By way of first non-limiting example,thermal properties of durable sheet materials of the present inventionmay be controlled, engineered, and/or tuned by controlling the size,number, and/or spatial distribution of air bubbles in the durable sheetmaterial. By way of second non-limiting example, thermal properties ofdurable sheet materials of the present invention may be controlled,engineered, and/or tuned by adding a thermal dopant having a desiredthermal property (e.g. heat capacity, thermal conductivity, thermaleffusivity, and combinations thereof) and thus modifying the samethermal property of the durable sheet material as a whole. By way ofthird non-limiting example, thermal properties of durable sheetmaterials of the present invention may be controlled, engineered, and/ortuned by controlling the mass, volume, thickness, spatial distribution,etc. of thermal dopants included in the durable sheet material, therebyproviding for an engineered or designed spatial pattern of heat exchangein and through the durable sheet material.

Embodiments of the present invention provide for the manufacture offungal textile materials, and particularly fungal leather analogs, fromintact cohesive fungal biomasses (e.g. fungal biomats produced bysurface fermentation or any other suitable process), size-reduced orhomogenized fungal biomasses, or any other physical form of fungalbiomass, especially filamentous fungal biomass. The materials of thepresent invention generally include both an inactivated fungal biomassand a component selected from the group consisting of a polymer, aplasticizer, a crosslinker, and a dye, and the methods of the presentinvention allow for the introduction of such component(s) to theinactivated fungal biomass to produce materials having desired chemical,physical, and/or thermal properties. The materials of the presentinvention may generally be provided as durable sheet materials suitablefor use in the same or similar applications as conventional textiles.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 2 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 3 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 4 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 5 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 6 is a generalized schematic of a method for making a fungaltextile material, according to embodiments of the present invention.

FIG. 7 is a graph of the tensile strength of an MK7 leather analogmaterial as a function of glycerol content, according to embodiments ofthe present invention.

FIG. 8 is a graph of the strain at break of an MK7 leather analogmaterial as a function of glycerol content, according to embodiments ofthe present invention.

FIG. 9 is a graph of the degree of swelling of an MK7 leather analogmaterial as a function of glycerol content, according to embodiments ofthe present invention.

FIG. 10 is a graph of the mass loss upon soaking of an MK7 leatheranalog material as a function of glycerol content, according toembodiments of the present invention.

FIG. 11 is a graph of the tensile strength of an MK7 leather analogmaterial as a function of loading ratio, according to embodiments of thepresent invention.

FIG. 12 is a graph of the strain at break of an MK7 leather analogmaterial as a function of loading ratio, according to embodiments of thepresent invention.

FIG. 13 is a graph of the degree of swelling of an MK7 leather analogmaterial as a function of loading ratio, according to embodiments of thepresent invention.

FIG. 14 is a graph of the mass loss upon soaking of an MK7 leatheranalog material as a function of loading ratio, according to embodimentsof the present invention.

FIG. 15 is a graph of the tensile strength of an MK7 leather analogmaterial as a function of polyvinyl alcohol:chitosan ratio, according toembodiments of the present invention.

FIG. 16 is a graph of the strain at break of an MK7 leather analogmaterial as a function of polyvinyl alcohol:chitosan ratio, according toembodiments of the present invention.

FIG. 17 is a graph of the degree of swelling of an MK7 leather analogmaterial as a function of polyvinyl alcohol:chitosan ratio, according toembodiments of the present invention.

FIG. 18 is a graph of the mass loss upon soaking of an MK7 leatheranalog material as a function of polyvinyl alcohol:chitosan ratio,according to embodiments of the present invention.

FIGS. 19A, 19B, 19C, and 19D are histograms of size-reduced fungalparticles after 10 seconds, 20 seconds, 40 seconds, and 60 seconds,respectively, of blending in a conventional household blender, accordingto embodiments of the present invention.

FIG. 20 is a graph of blend overrun, heating overrun, overall overrun,and density of solutions of fungal particles in water as a function ofloading ratio, according to embodiments of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, unless otherwise specified, the term “biodegradable”refers to a material that, under a given set of conditions (e.g. theconditions specified in ISO 20136:2017, “Leather—determination ofdegradability by micro-organisms”), biodegrades more quickly than “true”(i.e. animal) leather.

As used herein, unless otherwise specified, the term “degree ofswelling” refers to the relative amount of change in the mass of a soliditem when the solid is saturated with a liquid. By way of non-limitingexample, a solid item that has a mass of 200 g when dry and a mass of300 g when saturated with water has a degree of swelling in water of50%, or 0.5. Where the term “degree of swelling” is used herein withoutexplicitly identifying a liquid, the liquid may be assumed to be water.

As used herein, unless otherwise specified, the term “durable” refers toa material that has at least one of a tear strength of at least about 5N/mm, a tear force of at least about 5 N, and a tensile strength of atleast about 1.5 MPa.

As used herein, unless otherwise specified, the term “fungal biomass”refers to a mass of a fungus that has been cultured, fermented, or grownby any suitable process. It is to be expressly understood that fungalbiomass may be produced by any of a number of methods known in the artand disclosed herein, including but not limited to surface fermentationmethods, submerged fermentation methods, solid-substrate submergedfermentation (SSSF) methods, and methods as disclosed in PCT ApplicationPublication WO2019/099474 (“the '474 publication”), the entirety ofwhich is incorporated herein by reference.

As used herein, unless otherwise specified, the terms “hide leather” and“true leather” are interchangeable and each refer to a durable, flexiblematerial created by tanning the hide or skin of an animal.

As used herein, unless otherwise specified, the term “inactivated”refers to fungal biomass that has been killed or otherwise preventedfrom actively growing by a suitable inactivation means, e.g. boiling,steaming, rinsing, irradiating, freezing, treating with an aqueoussolution of at least 70% ethanol, treating with ethanol vapor, treatingwith bases or otherwise raising the pH (with or without heating),treating with acids or otherwise lowering the pH (with or withoutheating), or mechanically disrupting or destroying (such as by blendingor otherwise size-reducing). It is to be expressly understood that afungal biomass may be inactivated during, in combination with, and/or asa result of another process step, such as a size-reducing, liming, ordeliming step. As used herein, unless otherwise specified, the term“infiltration” refers to the permeation and/or saturation of a solutioninto a mass of solid material, such that the solution or a portionthereof is distributed in the mass of solid material, such as forexample and without limitation, a polymer solution permeating theinterstitial spaces in a fungal biomat comprised of mycelia. Withoutbeing bound by theory, the infiltration of a fungal mycelial biomasswith a solution comprising components such as polymers and plasticizers,results in a textile material having such components distributed in thebiomass after the solvent is removed by curing. Such a distribution canbe substantially uniformly distributed or not uniformly distributed.

As used herein, unless otherwise specified, the term “loading ratio”refers to a weight ratio of fungal biomass to polymer in a fungaltextile composition.

As used herein, unless otherwise specified, the term “mass loss uponsoaking” refers to the relative amount of mass lost by a solid itemafter soaking in a liquid, disregarding the mass of liquid absorbed bythe solid item. By way of non-limiting example, a solid item that has amass of 100 grams when dry and a mass (disregarding the mass of absorbedliquid) of 95 grams after soaking in water has a mass loss upon soakingin water of 5%. Where the term “mass loss upon soaking” is used hereinwithout explicitly identifying a liquid, the liquid may be assumed to bewater.

As used herein, unless otherwise specified, the term “sheet” refers to alayer of solid material having a generally flat or planar shape and ahigh ratio of surface area to thickness.

As used herein, unless otherwise specified, the term “tannin” refersgenerally to any molecule that forms strong bonds with proteinstructures, and more particularly to a molecule that, when applied tohide leather, bonds strongly to protein moieties within the collagenstructures of the skin to improve the strength and degradationresistance of the leather. The most commonly used types of tannins arevegetable tannins, i.e. tannins extracted from trees and plants, andchromium tannins such as chromium(III) sulfate. Other examples oftannins as that term is used herein include modified naturally derivedpolymers, biopolymers, and salts of metals other than chromium, e.g.aluminum silicate (sodium aluminum silicate, potassium aluminumsilicate, etc.).

Referring now to FIG. 1, one embodiment of a method 100 for making afungal textile material is illustrated. In a first step 110 of themethod 100 illustrated in FIG. 1, a fungal biomass is produced by any ofseveral suitable methods, including but not limited to methods describedin PCT Application PCT/US2017/020050, filed 28 Feb. 2017 (“the '050application”); PCT Application PCT/US2018/048626, filed 29 Aug. 2018(“the '626 application”); U.S. Provisional Patent Application62/811,421, filed 27 Feb. 2019 (“the '421 application”); and the '474publication, the entireties of all of which are hereby incorporated byreference. As described in the '050 application, the '626 application,and the '421 application, the fungal biomass may be grown by surfacefermentation in an artificial medium to form a cohesive structure ofinterwoven or interconnected mycelia called biomat. According to themethods described in the '050 application, the '626 application, and the'421 application, it may, in embodiments, be desirable to control an oilcontent and/or lipid content of the fungal biomass by providing a growthmedium having a preselected ratio of carbon to nitrogen. Particularly,the production of certain lipids or oils, or amounts thereof, by thefungal biomass may result in fungal textile materials having certaindesirable material characteristics, e.g. improved water resistance,decreased conditioning requirements, etc.; such characteristics may beamenable to control, engineering, or tuning by providing a preselectedmolar ratio of carbon to nitrogen in a fungal growth medium, which mayin embodiments be between about 5 and about 20, or between about 7 andabout 15. In some embodiments, the production of certain lipids or oilsby the fungal biomass, e.g. oleic acid, linoleic acid, eicosenoic acid,palmitic acid, stearic acid, arachidic acid, behenic acid, etc. mayallow for the use of certain polymers, solvents, etc. that may otherwisenot be suitable in the practice of the invention and thereby provideproperties of the fungal textile material that may otherwise beunattainable, or may provide additional or alternative synergisticeffects of this kind.

In an optional second step 120 of the method 100 illustrated in FIG. 1,the fungal biomass may be size-reduced by any suitable method, whichmay, by way of non-limiting example, comprise being processed (e.g. in ablender, food processor, or similar size-reducing device), compressed(e.g. by moving jaws, rolls, gyratory cones, or similar compressiondevice), impacted (e.g. by hammer, high-speed jet of material, rollers,or similar impact device), spray-dried, and the like. The size-reductionstep may be carried out in any suitable device (e.g. a blender) for anysuitable length of time (e.g. two minutes). During the size reductionstep, at least a portion of a cohesive interconnected or interwovenmycelial network of the fungal biomass may be disrupted or destroyed.

In a third step 130 of the method 100 illustrated in FIG. 1, the fungalbiomass is mixed with a solution of a synthetic polymer and/or abiopolymer. The synthetic polymer may be any synthetic polymer that issoluble in the solvent of choice, which may, but need not, be water; byway of non-limiting example, the synthetic polymer may be a polyvinylalcohol, a polyethylene glycol, a polysiloxane, a polyphosphazene, alow- and/or high-density polyethylene, a polypropylene, a polyvinylchloride, a polystyrene, a nylon, a polytetrafluoroethylene, athermoplastic polyurethane, a polychlorotrifluoroethylene, apolycaprolactone, a polyacrylic acid, and/or any one or more syntheticpolymers sold under various brand names (e.g. Bakelite, Kevlar, Mylar,Neoprene, Nomex, Orlon, Rilsan, Technora, Teflon, Twaron, Ultem,Vectran, Viton, Zylon, etc.). The biopolymer may be any polymericmolecule naturally produced by animals, plants, or fungi, including, byway of non-limiting example, cellulose, chitin, chitosan, collagen,fibroin, hyaluronic acid, keratin, alginates, starches, and combinationsthereof. In embodiments, the solution (or another solution with whichthe biomat is combined in the same step or a preceding or followingstep) may also comprise additional components, such as, by way ofnon-limiting example, a plasticizer (e.g. glycerol and esters thereof,polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g.mannitol, sorbitol) and esters thereof, epoxidized triglyceridevegetable oils (e.g. from soybean oil), castor oil, pentaerythritol,fatty acid esters, carboxylic ester-based plasticizers, trimellitates,adipates, sebacates, maleates, biological plasticizers, and combinationsand mixtures thereof etc.) and/or a crosslinker (e.g. homobifunctionalcrosslinkers, heterobifunctional crosslinkers, photoreactivecrosslinking agents, citric acid, tannic acid, suberic acid, adipicacid, succinic acid, extracted vegetable tannins, glyoxal, andcombinations thereof). It is to be expressly understood that thesize-reduction step (if any) and the mixing step can be carried outsimultaneously or sequentially in any order.

In a fourth step 140 of the method 100 illustrated in FIG. 1, thebiomass/solution mixture is stirred, typically at elevated temperature(by way of non-limiting example, about 90° C. to about 100° C.). Afterstirring, the biomass/solution mixture may optionally be further mixedwith a dye to provide a desired color to the fungal textile material. Insome embodiments, the dye may be added earlier in the process.

In a fifth step 150 of the method 100 illustrated in FIG. 1, thebiomat/solution mixture is cured, optionally after being cast into adesired shape. The curing step may involve drying or the initiation of achemical reaction and may drive off the solvent of the solution.

In a sixth step 160 of the method 100 illustrated in FIG. 1, the curedmaterial is heat-pressed to form the desired fungal textile material. Inembodiments, the fungal textile material may have at least one physical,mechanical, and/or aesthetic characteristic that mimics or closelyresembles a physical, mechanical, and/or aesthetic characteristic of aconventional textile material such as leather.

Certain embodiments of the methods of the present invention may omitsteps in which the fungal biomass is size-reduced (e.g. the second step120 illustrated in FIG. 1). In some such embodiments, the biomass (e.g.a biomat produced according to the methods described in the '050application, the '626 application, and/or the '421 application) may ormay not have been previously size-reduced. In other embodiments, thebiomass used may be a biomass that does not require size reduction, suchas a fungal paste produced by submerged fermentation methods as knownand described in the art.

Referring now to FIG. 2, another embodiment of a method 200 for making afungal textile material is illustrated. In a first step 210 of themethod 200 illustrated in FIG. 2, a fungal biomass is produced andprocessed by any of several suitable methods, including but not limitedto methods described in the '050 application, the '626 application, the'421 application, and the '474 publication. The biomass may be boiled,rinsed, irradiated, and/or pressed to inactivate the organism and/orremove excess water and/or other liquid. The biomass may also be frozen,particularly where it is desirable or necessary to store the biomass fora period of time prior to performing the later steps and thus to extendthe usable “shelf life” of the biomass.

In a second step 220 of the method 200 illustrated in FIG. 2, the fungalbiomass is thawed (if previously frozen); size-reduced by any suitablemethod, which may, by way of non-limiting example, comprise beingprocessed in a blender, food processor, mill, sonicator or similarsize-reducing device; and blended or otherwise homogenized with waterand, optionally, a pigment to provide a desired color to the fungaltextile material. The size-reduction sub-step may be carried out in anysuitable device (e.g. a blender) for any suitable length of time (e.g.two minutes). The blending/homogenizing sub-step produces a viscous,substantially homogeneous fungal paste. It is to be expressly understoodthat the size-reducing sub-step and the blending/homogenizing sub-stepmay be carried out simultaneously, sequentially in the same vessel, orsequentially in different vessels; by way of non-limiting example, waterand optionally a pigment may be added to a blender together with thefungal biomass prior to the size-reduction sub-step, and thesecomponents may be blended simultaneously in the blender, thus carryingout the size-reduction sub-step and the blending/homogenizing sub-stepsimultaneously in the same vessel. In some embodiments, size-reducingthe fungal biomass may also result in inactivation of the fungalbiomass, e.g. by disrupting cellular structure of the fungus.

In an optional third step 230 of the method 200 illustrated in FIG. 2,the viscous, substantially homogeneous paste is degassed by any suitablemethod, which may, by way of non-limiting example, comprise one or moreof agitation and vacuum treatment. Degassing the fungal material mayprovide improved qualities to the finished fungal textile product,including but not limited to a texture or “feel” that is moreaesthetically pleasing to a user and/or more similar to a replicatedmaterial (e.g. true leather). In some embodiments, the degassing may beomitted; particularly, it may, in some embodiments, be desirable toallow at least some air bubbles or pockets to remain in the fungalpaste, as this may impart certain desirable thermal or insulatingproperties to the finished fungal textile material.

In a fourth step 240 of the method 200 illustrated in FIG. 2, the fungalpaste is mixed with a solution of a polymer in a solvent of choice. Thesolvent may, but need not, be water. The polymer may, but need not, be abiopolymer, i.e. any polymeric molecule naturally produced by animals,plants, or fungi, including, by way of non-limiting example, cellulose,chitin, chitosan, collagen, fibroin, hyaluronic acid, keratin,alginates, starches, and combinations thereof. In embodiments, thesolution (or another solution with which the biomat is combined in thesame step or a preceding or following step) may comprise, in addition toor as an alternative to a biopolymer, a synthetic polymer soluble in thesolvent (e.g. a polyvinyl alcohol, a polyethylene glycol, apolysiloxane, a polyphosphazene, a low- and/or high-densitypolyethylene, a polypropylene, a polyvinyl chloride, a polystyrene, anylon, a polytetrafluoroethylene, a thermoplastic polyurethane, apolychlorotrifluoroethylene, a polycaprolactone, a polyacrylic acid,and/or any one or more synthetic polymers sold under various brand names(e.g. Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora,Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). In furtherembodiments, the solution may comprise one or more additionalcomponents, such as a plasticizer (e.g. glycerol and esters thereof,polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g.mannitol, sorbitol) and esters thereof, epoxidized triglyceridevegetable oils (e.g. from soybean oil), castor oil, pentaerythritol,fatty acid esters, carboxylic ester-based plasticizers, trimellitates,adipates, sebacates, maleates, biological plasticizers, and combinationsand mixtures thereof etc.), a crosslinker (e.g. homobifunctionalcrosslinkers, heterobifunctional crosslinkers, photoreactivecrosslinking agents, citric acid, tannic acid, suberic acid, adipicacid, succinic acid, extracted vegetable tannins, glyoxal, andcombinations thereof), a solubilizer (e.g. hydrochloric acid, aceticacid, formic acid, lactic acid, etc.), and/or a pH adjusting agent (e.g.hydrochloric acid, acetic acid, formic acid, lactic acid, etc.).

The solution may be made by combining the polymer and the solvent, andoptionally one or more additional components, in a vessel and heatingthe combination while stirring. In embodiments in which the solutionincludes a solubilizer and/or a pH adjusting agent, either or both ofthese may be added to the solution after heating and stirring of theother components. Preferably, the polymer (biopolymer, syntheticpolymer, or a combination thereof) is completely dissolved in thesolvent before the solution is mixed with the optionally degassed fungalpaste. The mixture may be heated (e.g. to about 90° C. and/or toboiling) and/or stirred for a time sufficient to ensure that the mixtureis substantially homogeneous, e.g. between about 30 minutes and about 45minutes.

In an optional fifth step 250 of the method 200 illustrated in FIG. 2,the mixture produced in the fourth step is degassed by any suitablemethod, which may, by way of non-limiting example, comprise one or moreof agitation and vacuum treatment. Degassing the mixture may provideimproved qualities to the finished fungal textile product, including butnot limited to a texture or “feel” that is more aesthetically pleasingto a user and/or more similar to a replicated material (e.g. trueleather). In some embodiments, the degassing may be omitted;particularly, it may, in some embodiments, be desirable to allow atleast some air bubbles or pockets to remain in the mixture, as this mayimpart certain desirable thermal or insulating properties to thefinished fungal textile material.

In a sixth step 260 of the method 200 illustrated in FIG. 2, the fungalmixture is cured, optionally after being cast into a desired shape (e.g.a flat or textured mold). The curing step may or may not involve curingor the initiation of a chemical reaction and may or may not drive offthe solvent of the solution. The curing step may be carried out underambient air at room temperature. The curing may be allowed to continueunder conditions for a time sufficient to provide a desired mass (e.g.about 20% of the mass prior to drying/curing) and/or moisture content ofthe cured material.

In an optional seventh step 270 of the method 200 illustrated in FIG. 2,the cured material may be heat-pressed to form the desired fungaltextile material. In embodiments, the fungal textile material may haveat least one physical, mechanical, and/or aesthetic characteristic thatmimics or closely resembles a physical, mechanical, and/or aestheticcharacteristic of a conventional textile material such as leather. Thetemperature (e.g. about 100° C.) and/or time (e.g. between about 10minutes and about 20 minutes) of the heat-pressing may be selected toprovide the desired physical, mechanical, and/or aestheticcharacteristic. The fungal textile material may, but need not, then belaminated to a textile backing; in these embodiments, a portion of thesolution of the fourth step may, but need not, be utilized as anadhesive for adhering the fungal textile material to the textilebacking.

Generally, the methods illustrated in FIGS. 1 and 2 cause a network offungal filaments to be crosslinked together by a combination of apolymer (e.g. chitosan) and a crosslinker (e.g. citric acid). Thepolymer and crosslinker can form bonds via esterification reactions(between alcohol groups of the fungal filaments and/or the polymer, andcarboxylic acid groups of the crosslinker and/or the fungal filaments)and/or amidation reactions (between amide groups of the fungal filamentsand/or the polymer, and carboxylic acid groups of the crosslinker and/orthe fungal filaments). These reactions may be catalyzed by, e.g. acidicconditions and/or heat (e.g. in a heat-pressing step). The use of aplasticizer such as glycerol can impart flexibility to the finishedfungal textile material. The method of FIG. 1 can be used in conjunctionwith both intact and size-reduced fungal biomasses.

Referring now to FIG. 3, another embodiment of a method 300 for making afungal textile material is illustrated. In a liming step 310 of themethod 300 illustrated in FIG. 3, an inactivated fungal biomass is addedto an aqueous mixture or solution of components and agitated, e.g. on ashaker table. The aqueous mixture or solution comprises an aqueoussolvent, a mass of which is typically about equal to that of the fungalbiomass, and a liming substance, most commonly calcium hydroxide (i.e.slaked lime), in an amount of between about 0.01 wt % and about 6 wt %or any sub-range between those values, most commonly about 3 wt %,relative to the weight of the fungal biomass. The aqueous mixture orsolution may optionally further include a solubilizer or surfactant,such as a polysorbate, in an amount of between about 0.01 wt % and about1 wt % or any sub-range between those values, most commonly about 0.2 wt%, relative to the weight of the fungal biomass. The agitation may becarried out for any suitable time between about 1 minute and about 180minutes or any sub-range between those values, most commonly about 90minutes.

Prior to step 310, the fungal biomass may have been produced andprocessed by any of several suitable methods, including but not limitedto methods described in the '050 application, the '626 application, the'421 application, and the '474 publication, and may be boiled, rinsed,irradiated, and/or pressed to inactivate the organism and/or removeexcess water and/or other liquid. The biomass may also, prior to step310, be frozen, particularly where it is desirable or necessary to storethe biomass for a period of time prior to performing the later steps andthus to extend the usable “shelf life” of the biomass, and subsequentlythawed.

Step 310 of the method 300 illustrated in FIG. 3 may be carried out onan intact fungal biomass, e.g. a cohesive fungal biomat produced bysurface fermentation, or it may be carried out on a fungal biomass thathas previously been size-reduced by any suitable method, which may, byway of non-limiting example, comprise being processed in a blender, foodprocessor, mill, sonicator or similar size-reducing device. Any suchsize reduction may be carried out in any suitable device (e.g. ablender) for any suitable length of time (e.g. two minutes). In someembodiments, the fungal biomass may be active prior to size reductionand may be inactivated as a result of the size reduction, e.g. bydisrupting cellular structure of the fungus. More generally, it is to beexpressly understood that the fungal biomass may be inactivated during,in combination with, or as a result of any one or more other steps ofthe method 300, e.g. the liming step 310 (in which the pH of the fungalbiomass is raised to at least about 7, or another pH sufficiently highto kill the fungus) or any of the other steps that follow (particularlyif carried out at elevated temperature).

In a deliming step 320 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass is added to an aqueous mixture or solution ofcomponents and agitated, e.g. on a shaker table. The aqueous mixture orsolution comprises an aqueous solvent, a mass of which is typicallyabout half that of the mass of the starting (i.e. prior to step 310)fungal biomass, and a deliming substance, most commonly ammoniumsulfate, in an amount of between about 0.01 wt % and about 6 wt % or anysub-range between those values, most commonly about 3 wt %, relative tothe weight of the starting (i.e. prior to step 310) fungal biomass. Theaqueous mixture or solution may optionally further include a solubilizeror surfactant, such as a polysorbate, in an amount of between about 0.01wt % and about 1 wt % or any sub-range between those values, mostcommonly about 0.2 wt %, relative to the weight of the starting (i.e.prior to step 310) fungal biomass. The agitation may be carried out forany suitable time between about 1 minute and about 180 minutes or anysub-range between those values, most commonly about 90 minutes.

In a pickling step 330 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass is mixed with a solution of a polymer in anaqueous solvent. The polymer may, but need not, be a biopolymer, i.e.any polymeric molecule naturally produced by animals, plants, or fungi,including, by way of non-limiting example, cellulose, chitin, chitosan,collagen, fibroin, hyaluronic acid, keratin, alginates, starches, andcombinations thereof. In embodiments, the solution (or another solutionwith which the inactivated fungal biomass is combined in the same stepor a preceding or following step) may comprise, in addition to or as analternative to a biopolymer, a synthetic polymer soluble in the solvent(e.g. a polyvinyl alcohol, a polyethylene glycol, a polysiloxane, apolyphosphazene, a low- and/or high-density polyethylene, apolypropylene, a polyvinyl chloride, a polystyrene, a nylon, apolytetrafluoroethylene, a thermoplastic polyurethane, apolychlorotrifluoroethylene, a polycaprolactone, a polyacrylic acid,and/or any one or more synthetic polymers sold under various brand names(e.g. Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora,Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). In furtherembodiments, the solution may comprise one or more additionalcomponents, such as a plasticizer (e.g. glycerol and esters thereof,polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g.mannitol, sorbitol) and esters thereof, epoxidized triglyceridevegetable oils (e.g. from soybean oil), castor oil, pentaerythritol,fatty acid esters, carboxylic ester-based plasticizers, trimellitates,adipates, sebacates, maleates, biological plasticizers, and combinationsthereof), a crosslinker (e.g. homobifunctional crosslinkers,heterobifunctional crosslinkers, photoreactive crosslinking agents,citric acid, tannic acid, suberic acid, adipic acid, succinic acid,extracted vegetable tannins, glyoxal, and combinations thereof), asolubilizer (e.g. hydrochloric acid, acetic acid, formic acid, lacticacid, etc.), and/or a pH adjusting agent (e.g.

hydrochloric acid, acetic acid, formic acid, lactic acid, etc.). Analkali metal halide (e.g. sodium chloride) may be provided to preventswelling of the inactivated fungal biomass.

The solution may be made by combining the polymer and the solvent, andoptionally one or more additional components, in a vessel and agitatingor stirring the combination, optionally while heating the combination.In embodiments in which the solution includes a solubilizer and/or a pHadjusting agent, either or both of these may be added to the solutionafter heating and stirring of the other components. Preferably, thepolymer (biopolymer, synthetic polymer, or a combination thereof) iscompletely dissolved in the solvent before the solution is mixed withthe optionally degassed fungal paste. The mixture may be heated (e.g. toabout 90° C. and/or to boiling) and/or stirred for a time sufficient toensure that the mixture is substantially homogeneous, e.g. between about1 minute and about 240 minutes or any sub-range between those values,and most typically between about 30 minutes and about 45 minutes orabout 120 minutes.

The polymer solution to which the inactivated fungal biomass is added instep 330 of the method 300 generally includes a mass of aqueous solventthat is generally about equal to that of the starting (i.e. prior tostep 310) fungal biomass; and the polymer in an amount of between about0.01 wt % and about 10 wt % or any sub-range between those values, mostcommonly about 1 wt %, relative to the starting (i.e. prior to step 310)fungal biomass. Other components, if present during step 330, may beprovided in any appropriate amount; by way of non-limiting example, asolubilizer or a pH adjusting agent may be provided in an amount ofbetween about 0.01 wt % and about 10 wt % or any sub-range between thosevalues, most commonly between about 0.5 wt % and about 2.5 wt %,relative to the starting (i.e. prior to step 310) fungal biomass, andthe alkali metal halide may be provided in an amount of between about0.01 wt % and about 14 wt % or any sub-range between those values, mostcommonly about 7 wt %, relative to the starting (i.e. prior to step 310)fungal biomass.

In a tanning step 340 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass from the pickling step 330 is added to anaqueous solution comprising a crosslinking or tanning agent andagitated, e.g. on a shaker table. The aqueous solution comprises anaqueous solvent, a mass of which is typically about equal to that of themass of the starting (i.e. prior to step 310) fungal biomass, and acrosslinking or tanning agent, e.g. citric acid and/or tannic acid, inan amount of between about 0.01 wt % and about 12 wt %, most commonlyabout 5 wt %, relative to the weight of the starting (i.e. prior to step310) fungal biomass. The agitation may be carried out for any suitabletime between about 1 minute and about 360 minutes or any sub-rangebetween those values, most commonly about 180 minutes.

Although not illustrated in FIG. 3, the method 300 may optionallycomprise one or more rinsing steps, in which the inactivated fungalbiomass is rinsed with water to remove excess aqueous solution, afterany one or more of liming step 310, deliming step 320, pickling step330, and tanning step 340. A rinsing step may comprise draining thevessel containing the inactivated fungal biomass (e.g. a shaker flask)of excess aqueous solution, refilling the vessel with water, agitatingthe vessel, and draining the vessel of water.

In a plasticizing step 350 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass is added to an aqueous solution comprising aplasticizer and agitated, e.g. on a shaker table. The aqueous solutioncomprises an aqueous solvent, a mass of which is typically about equalto that of the mass of the starting (i.e. prior to step 310) fungalbiomass, and a plasticizer, e.g. glycerol, in an amount of between about0.01 wt % and about 50 wt % or any sub-range between those values, mostcommonly about 25 wt %, relative to the weight of the starting (i.e.prior to step 310) fungal biomass. The agitation may be carried out forany suitable time between about 1 minute and about 180 minutes or anysub-range between those values, most commonly about 90 minutes. In someembodiments, the plasticizing step 350 may be a fatliquoring step, i.e.the plasticizer may be a fatliquoring oil such as sulfated castor oil,beeswax, coconut oil, vegetable oil, olive oil, linseed oil, oleic acid,sulfated fish oil, sulfated canola oil, soybean oil, palm oil, fattyacids, or a combination thereof.

In a drying step 360 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass is dried, optionally after being cast into adesired shape (e.g. a flat or textured mold) if produced from asize-reduced fungal biomass. The drying step may or may not involve theinitiation of a chemical reaction, but generally results in at leastmost of any residual water, solvents, and other liquids being drivenfrom the inactivated fungal biomass. The drying may be passive (i.e. atroom temperature without the use of a blower, fan, etc.) or active (i.e.under heating and/or using forced air, dry milling, etc.); when thedrying is active, the temperature may be raised to a desired temperatureabove room temperature, most commonly about 80° F., and/or any suitableair forcing means (e.g. a blower, a fan, a forced-air dehydrator, etc.)may be used. In some embodiments, at least a portion of the fungalmaterial may be clamped or otherwise pressed to reduce shrinkage. Thecuring may be allowed to continue under conditions for a time sufficientto provide a desired mass (e.g. about 20% of the mass prior todrying/curing) and/or moisture content of the cured material, which mayin embodiments be between about 1 minute and about 2 days or anysub-range between those values, most commonly about 1 day.

In a heat-pressing step 370 of the method 300 illustrated in FIG. 3, theinactivated fungal biomass is heat-pressed to form the desired fungaltextile material. In embodiments, the fungal textile material may haveat least one physical, mechanical, and/or aesthetic characteristic thatmimics or closely resembles a physical, mechanical, and/or aestheticcharacteristic of a conventional textile material such as leather;particularly, the heat-pressing step may be configured to impart aleather-like texture to the fungal textile material. The temperature(e.g. about 100° C.) and/or time (e.g. between about 1 minute and about20 minutes, most commonly about 10 minutes) of the heat-pressing may beselected to provide the desired physical, mechanical, and/or aestheticcharacteristic. The fungal textile material may, but need not, then belaminated to a non-fungal textile backing.

Referring now to FIG. 4, another embodiment of a method 400 for making afungal textile material is illustrated. In an inactivating step 405, thefungal biomass is inactivated to prevent active growth and metabolism ofthe fungus. This inactivation may commonly be effected by boiling thefungal biomass in a sufficient volume of water to completely submerge orsurround the fungal biomass; this boiling is typically conducted for aperiod of between about 1 minute and about 60 minutes or any sub-rangebetween those values, most commonly about 30 minutes. Of course, theinactivating step 405 may also be conducted by any other suitable means,such as by irradiating, freezing, size-reducing, or a combination ofthese with or without boiling.

Prior to step 405, the fungal biomass may have been produced andprocessed by any of several suitable methods, including but not limitedto methods described in the '050 application, the '626 application, the'421 application, and the '474 publication. The biomass may also, priorto step 405, be frozen, particularly where it is desirable or necessaryto store the biomass for a period of time prior to performing the latersteps and thus to extend the usable “shelf life” of the biomass, andsubsequently thawed.

Step 405 of the method 400 illustrated in FIG. 4 may be carried out onan intact fungal biomass, e.g. a cohesive fungal biomat produced bysurface fermentation, or it may be carried out on a fungal biomass thathas previously been size-reduced by any suitable method, which may, byway of non-limiting example, comprise being processed in a blender, foodprocessor, mill, sonicator or similar size-reducing device. Any suchsize reduction may be carried out in any suitable device (e.g. ablender) for any suitable length of time (e.g. two minutes). In someembodiments, the fungal biomass may be active prior to size reductionand may be inactivated as a result of the size reduction, e.g. bydisrupting cellular structure of the fungus.

Step 405 generally also includes dissolving, mixing, or suspending theinactivated fungal biomass in an aqueous solvent and may also includeadding a solubilizer or surfactant, e.g. a polysorbate, to theinactivated fungal biomass and combining the solubilizer or surfactantwith the inactivated fungal biomass, e.g. by agitation. A mass of theaqueous solvent may generally be between about half and about six times,most commonly about three times, that of the inactivated fungal biomass.The solubilizer or surfactant may be provided in an amount of betweenabout 0.01 wt % and about 1 wt % or any sub-range between those values,most commonly about 0.2 wt %, relative to the weight of the fungalbiomass. The agitation or other mechanical manipulation to combine theinactivated fungal biomass with the aqueous solvent, and optionally thesolubilizer or surfactant, may be carried out for a period of betweenabout 1 minute and about 60 minutes or any sub-range between thosevalues, most commonly about 30 minutes.

In a liming step 415 of the method 400 illustrated in FIG. 4, theinactivated fungal biomass is added to an aqueous mixture or solution ofcomponents and agitated, e.g. on a shaker table. The aqueous mixture orsolution comprises an aqueous solvent, a mass of which is typicallyabout equal to that of the fungal biomass, and a liming substance, mostcommonly calcium hydroxide (i.e. slaked lime), in an amount of betweenabout 0.01 wt % and about 10 wt % or any sub-range between those values,most commonly about 3 wt %, relative to the weight of the fungalbiomass. The aqueous mixture or solution may optionally further includea solubilizer or surfactant, such as a polysorbate, in an amount ofbetween about 0.01 wt % and about 1 wt % or any sub-range between thosevalues, most commonly about 0.2 wt %, relative to the weight of thefungal biomass. The agitation may be carried out for any suitable timebetween about 1 minute and about 300 minutes or any sub-range betweenthose values, most commonly about 150 minutes.

In a deliming step 425 of the method 400 illustrated in FIG. 4, theinactivated fungal biomass is added to an aqueous mixture or solution ofcomponents and agitated, e.g. on a shaker table. The aqueous mixture orsolution comprises an aqueous solvent, a mass of which is typicallyabout half that of the mass of the starting (i.e. prior to step 405)fungal biomass, and a deliming substance, most commonly ammonium sulfateor ammonium chloride, in an amount of between about 0.01 wt % and about10 wt % or any sub-range between those values, most commonly about 3 wt%, relative to the weight of the starting (i.e. prior to step 405)fungal biomass. The aqueous mixture or solution may optionally furtherinclude a solubilizer or surfactant, such as a polysorbate, in an amountof between about 0.01 wt % and about 0.4 wt % or any sub-range betweenthose values, most commonly about 0.2 wt %, relative to the weight ofthe starting (i.e. prior to step 405) fungal biomass. The agitation maybe carried out for any suitable time between about 1 minute and about150 minutes or any sub-range between those values, most commonly about75 minutes.

In a pickling step 435 of the method 400 illustrated in FIG. 4, theinactivated fungal biomass is mixed with an acid, most commonlyhydrochloric acid, or other pH adjusting agent. Sufficient pH adjustingagent is added to achieve a target pH of no more than about 4.0,typically between about 0.5 and about 3.5, more typically between about1.0 and about 3.0, even more typically between about 1.5 and about 2.5,and most typically about 2.0. It is generally desirable to choose amolarity and/or molality of the acid, or a concentration of pH adjustingagent in an aqueous solvent, that allows this target pH to be achievedby adding a preselected mass or volume of acid or liquid solution. Theacid or aqueous solution of pH adjusting agent may further comprise analkali metal halide, e.g. sodium chloride, to prevent swelling of thefungal biomass; the alkali metal halide may be present in an amount ofbetween about 0.01 wt % and about 14 wt % or any sub-range between thosevalues, most commonly about 7 wt %, relative to the starting (i.e. priorto step 405) fungal biomass. The inactivated fungal biomass may beagitated together with the acid and/or pH adjusting agent, andoptionally the alkali metal halide, for a period of between about 1minute and about 180 minutes or any sub-range between those values, mostcommonly about 90 minutes.

In a tanning step 445 of the method 400 illustrated in FIG. 4, a firstcrosslinking or tanning agent is added to the inactivated fungal biomassand the combination is agitated, e.g. in a drum or on a shaker table.The crosslinking or tanning agent may in embodiments comprise analdehyde, an aluminum salt, a chromium salt, or a titanium salt, and maycommonly comprise an aluminum silicate. The crosslinking or tanningagent may generally be provided in an amount of between about 0.01 wt %and about 15 wt % or any sub-range between those values, most commonlybetween about 1.5 wt % and about 7.5 wt %, relative to the weight of thestarting (i.e. prior to step 405) fungal biomass. The agitation may becarried out for any suitable time between about 1 minute and about 180minutes or any sub-range between those values, most commonly betweenabout 30 minutes and about 150 minutes. During the agitation, a base orother pH adjusting agent, e.g. sodium hydroxide, may commonly be added,either at one time or at multiple times, to achieve and/or maintain atarget pH, which in embodiments is generally between about 2.0 and about6.0, typically between about 2.5 and about 5.5, more typically betweenabout 3.0 and about 5.0, even more typically between about 3.5 and about4.5, and most typically about 4.0.

Although not illustrated in FIG. 4, the method 400 may optionallycomprise one or more rinsing steps, in which the inactivated fungalbiomass is rinsed with water to remove excess aqueous solution, afterany one or more of liming step 415, deliming step 425, and tanning step445. A rinsing step may comprise draining the vessel containing theinactivated fungal biomass (e.g. a shaker flask) of excess aqueoussolution, refilling the vessel with water, agitating the vessel, anddraining the vessel of water.

In a re-tanning step 455 of the method 400 illustrated in FIG. 4, asecond crosslinking or tanning agent is added to the inactivated fungalbiomass and the combination is agitated, e.g. in a drum or on a shakertable. The second crosslinking or tanning agent may in embodimentscomprise, e.g., citric acid, and may be provided in an amount of betweenabout 0.01 wt % and about 6 wt % or any sub-range between those values,most commonly about 3 wt %, relative to the weight of the starting (i.e.prior to step 410) fungal biomass. The agitation may be carried out forany suitable time between about 1 minute and about 480 minutes or anysub-range between those values, most commonly about 60 minutes.

The re-tanning step 455 may optionally comprise additional substeps toimpart additional substances or characteristics to the inactivatedfungal biomass and thus to the finished fungal textile material. By wayof first non-limiting example, the inactivated fungal biomass may bemixed with an aqueous solution of any polymer as disclosed herein andagitated, e.g. in a drum or on a shaker table. The polymer may beprovided in an amount of between about 0.01 wt % and about 30 wt % orany sub-range between those values, most commonly between about 0.5 wt %and about 5 wt %, relative to the weight of the starting (i.e. prior tostep 410) fungal biomass. The agitation may be carried out for anysuitable time between about 1 minute and about 480 minutes or anysub-range between those values, most commonly about 60 minutes. By wayof second non-limiting example, a dye, such as an anionic dye, may beadded to the inactivated fungal biomass and the combination may beagitated, e.g. in a drum or on a shaker table, for a time sufficient toimpart a desired color to the inactivated fungal biomass (typicallybetween about 1 minute and about 240 minutes or any sub-range betweenthose values, and most typically about 120 minutes). The addition of theoptional components (e.g. polymer, dye, etc.) may be carried out before,after, or simultaneously with addition of the second crosslinking ortanning agent.

Throughout the re-tanning step 455, acids, bases, and/or other pHadjusting agents may be added to maintain a target pH. By way of firstnon-limiting example, it may, in some embodiments, be desirable to beginthe re-tanning step 455 at an initial pH of between about 2.0 and about6.0 (more typically between about 2.5 and about 5.5, more typicallybetween about 3.0 and about 5.0, more typically between about 3.5 andabout 4.5, and most typically about 4.0) and gradually raise the pH tobetween about 3.5 and about 7.5 (more typically between about 4.0 andabout 7.0, more typically between about 4.5 and about 6.5, moretypically between about 5.0 and about 6.0, and most typically about 5.5)by adding a base or other pH increasing agent in one or more aliquotsduring agitation. By way of second non-limiting example, where there-tanning step 455 includes the addition of a polymer, it may, in someembodiments, be desirable to maintain a pH of between about 3.5 andabout 7.5 (more typically between about 4.0 and about 7.0, moretypically between about 4.5 and about 6.5, more typically between about5.0 and about 6.0, and most typically about 5.5) during agitation of theinactivated fungal biomass together with the polymer.

The polymer may, but need not, be a biopolymer, i.e. any polymericmolecule naturally produced by animals, plants, or fungi, including, byway of non-limiting example, cellulose, chitin, chitosan, collagen,fibroin, hyaluronic acid, keratin, alginates, starches, and combinationsthereof. In embodiments, the solution (or another solution with whichthe inactivated fungal biomass is combined in the same step or apreceding or following step) may comprise, in addition to or as analternative to a biopolymer, a synthetic polymer soluble in the solvent(e.g. a polyvinyl alcohol, a polyethylene glycol, a polysiloxane, apolyphosphazene, a low- and/or high-density polyethylene, apolypropylene, a polyvinyl chloride, a polystyrene, a nylon, apolytetrafluoroethylene, a thermoplastic polyurethane, apolychlorotrifluoroethylene, a polycaprolactone, a polyacrylic acid,and/or any one or more synthetic polymers sold under various brand names(e.g. Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora,Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). In furtherembodiments, the solution may comprise one or more additionalcomponents, such as a plasticizer (e.g. glycerol and esters thereof,polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g.mannitol, sorbitol) and esters thereof, epoxidized triglyceridevegetable oils (e.g. from soybean oil), castor oil, pentaerythritol,fatty acid esters, carboxylic ester-based plasticizers, trimellitates,adipates, sebacates, maleates, biological plasticizers, and combinationsand mixtures thereof etc.), a crosslinker (e.g. homobifunctionalcrosslinkers, heterobifunctional crosslinkers, photoreactivecrosslinking agents, citric acid, tannic acid, suberic acid, adipicacid, succinic acid, extracted vegetable tannins, glyoxal, andcombinations thereof), a solubilizer (e.g. hydrochloric acid, aceticacid, formic acid, lactic acid, etc.), and/or a pH adjusting agent (e.g.hydrochloric acid, acetic acid, formic acid, lactic acid, etc.). Analkali metal halide (e.g. sodium chloride) may be provided to preventswelling of the inactivated fungal biomass.

The solution may be made by combining the polymer and the solvent, andoptionally one or more additional components, in a vessel and agitatingor stirring the combination, optionally while heating the combination.In embodiments in which the solution includes a solubilizer and/or a pHadjusting agent, either or both of these may be added to the solutionafter heating and stirring of the other components. Preferably, thepolymer (biopolymer, synthetic polymer, or a combination thereof) iscompletely dissolved in the solvent before the solution is mixed withthe inactivated fungal biomass. The mixture may be heated (e.g. to about90° C. and/or to boiling) and/or stirred for a time sufficient to ensurethat the mixture is substantially homogeneous, e.g. between about 1minute and about 240 minutes, and most typically between about 30minutes and about 45 minutes or about 120 minutes.

In a plasticizing step 465 of the method 400 illustrated in FIG. 4, aplasticizer is added to the inactivated fungal biomass and thecombination is agitated, e.g. in a drum or on a shaker table. Inembodiments, the plasticizing step may be a fatliquoring step, i.e. theplasticizer may comprise a fatliquoring oil such as sulfated castor oil,beeswax, coconut oil, vegetable oil, olive oil, linseed oil, oleic acid,sulfated fish oil, sulfated canola oil, soybean oil, palm oil, fattyacids, or a combination thereof, and may be provided in any suitableamount. The agitation may be carried out for any suitable time betweenabout 1 minute and about 120 minutes or any sub-range between thosevalues, most commonly about 60 minutes. The plasticizer may be providedas an emulsion, especially when the plasticizer is a traditional leatherfatliquoring oil, and in some such embodiments the plasticizing step 465may be concluded by adding an acid, e.g. hydrochloric acid, to theemulsion to split the emulsion and allow for easier draining and removalof the plasticizer.

In a backing step 475 of the method 400 illustrated in FIG. 4, at leastone backing layer of a non-fungal textile material is applied to theinactivated fungal biomass and adhered to the inactivated fungalbiomass. The non-fungal textile material may, in embodiments, includeany one or more of an acrylic textile, an alpaca textile, an angoratextile, a cashmere textile, a coir textile, a cotton textile, aneisengarn textile, a hemp textile, a jute textile, a Kevlar textile, alinen textile, a microfiber textile, a mohair textile, a nylon textile,an olefin textile, a pashmina textile, a polyester textile, a piñatextile, a ramie textile, a rayon textile, a sea silk textile, a silktextile, a sisal textile, a spandex textile, a spider silk textile, anda wool textile. The adhesive may be any suitable laminating adhesiveused in textiles, e.g. polyvinyl acetate, and may in some embodimentsinclude any suitable amount of a crosslinker or plasticizer, e.g. citricacid.

In a heat-pressing step 485 of the method 400 illustrated in FIG. 4, theinactivated fungal biomass, together with the non-fungal textilebacking, is heat-pressed. In embodiments, the fungal textile materialmay have at least one physical, mechanical, and/or aestheticcharacteristic that mimics or closely resembles a physical, mechanical,and/or aesthetic characteristic of a conventional textile material suchas leather; particularly, the heat-pressing step may be configured toimpart a leather-like texture to the fungal textile material. Thetemperature (e.g. about 100° C.) and/or time (e.g. between about 1minute and about 20 minutes, most commonly about 10 minutes) of theheat-pressing may be selected to provide the desired physical,mechanical, and/or aesthetic characteristic.

In a drying step 495 of the method 400 illustrated in FIG. 4, theinactivated fungal biomass is dried, optionally after being cast into adesired shape (e.g. a flat or textured mold), to form the fungal textilematerial. The drying step may or may not involve the initiation of achemical reaction, but generally results in at least most of anyresidual water, solvents, and other liquids being driven from theinactivated fungal biomass. The drying may be passive (i.e. at roomtemperature without the use of a blower, fan, etc.) or active (i.e.under heating and/or using forced air); when the drying is active, thetemperature may be raised to a desired temperature above roomtemperature, most commonly about 80° F., and/or any suitable air forcingmeans (e.g. a blower, a fan, a forced-air dehydrator, etc.) may be used.In some embodiments, at least a portion of the fungal material may beclamped or otherwise pressed to reduce shrinkage. The curing may beallowed to continue under conditions for a time sufficient to provide adesired mass (e.g. about 18% of the mass prior to drying/curing) and/ormoisture content of the cured material, which may in embodiments bebetween about 1 minute and about 2 days, most commonly about 1 day.

Although not illustrated in FIG. 4, the method 400 may include at leastone additional post-processing or final handling step. Particularly, oneor more traditional leather finishing waxes or oils (e.g. carnauba wax,candelilla wax) or nitrocellulose may be added to the fungal textilematerial, in any suitable amount and for any suitable time. Referringnow to FIG. 5, another embodiment of a method 500 for making a fungaltextile material is illustrated. In an inactivating step 510 of themethod 500 illustrated in FIG. 5, a fungal biomass is inactivated asdescribed herein, e.g. with respect to inactivating step 405 of themethod 400 illustrated in FIG. 4. In a liming step 520 of the method 500illustrated in FIG. 5, the inactivated fungal biomass is limed asdescribed herein, e.g. with respect to liming step 310 of the method 300illustrated in FIG. 3 and/or liming step 415 of the method 400illustrated in FIG. 4. In a deliming step 530 of the method 500illustrated in FIG. 5, the inactivated fungal biomass is delimed asdescribed herein, e.g. with respect to deliming step 320 of the method300 illustrated in FIG. 3 and/or deliming step 425 of the method 400illustrated in FIG. 4.

In a pickling step 540 of the method 500 illustrated in FIG. 5, theinactivated fungal biomass is pickled as described herein, e.g. withrespect to pickling step 330 of the method 300 illustrated in FIG. 3and/or pickling step 435 of the method 400 illustrated in FIG. 4.However, one difference in the pickling step 540 of the method 500illustrated in FIG. 5 relative to the pickling steps of otherembodiments lies in the addition of at least two aliquots ofcrosslinker, e.g. tannic acid, to the combination of inactivated fungalbiomass and polymer solution, or vice versa, such that the inactivatedfungal biomass may be contacted with the polymer solution before beingcontacted with the first aliquot of crosslinker, or simultaneously withbeing contacted with the first aliquot of crosslinker, or after beingcontacted with the first aliquot of crosslinker but before beingcontacted with the second aliquot of crosslinker, or simultaneously withbeing contacted with the second aliquot of crosslinker, or after beingcontacted with the second aliquot of crosslinker. In this way, themethod 500 of FIG. 5 may, in a sense, combine pickling, tanning, andre-tanning steps, e.g. steps 330 and 340 of method 300 and/or steps 435,445, and 455 of method 400, into a single step comprising pickling,tanning, and re-tanning substeps.

In a neutralizing step 550 of the method 500 illustrated in FIG. 5, thepH of the inactivated fungal biomass is neutralized by contacting theinactivated fungal biomass with a pH neutralizing agent, which in mostembodiments is a basic pH neutralizing agent, e.g. sodium bicarbonate,but may in some embodiments be an acidic pH neutralizing agent. The pHneutralizing agent may be provided as part of an aqueous solution, andmay (but need not) be provided in a suitable amount to provide a pH ofabout 7. As with other steps, the neutralizing step 550 may be carriedout with agitation, e.g. in a shaker flask.

In a plasticizing step 560 of the method 500 illustrated in FIG. 5, theinactivated fungal biomass is plasticized as described herein, e.g. withrespect to plasticizing step 350 of the method 300 illustrated in FIG. 3and/or plasticizing step 465 of the method 400 illustrated in FIG. 4. Ina heat-pressing step 570 of the method 500 illustrated in FIG. 5, theinactivated fungal biomass is heat-pressed as described herein, e.g.with respect to heat-pressing step 370 of the method 300 illustrated inFIG. 3 and/or heat-pressing step 485 of the method 400 illustrated inFIG. 4.

Referring now to FIG. 6, another embodiment of a method 600 for making afungal textile material is illustrated. In an inactivating step 610 ofthe method 600 illustrated in FIG. 6, a fungal biomass is inactivated asdescribed herein, e.g. with respect to inactivating step 405 of themethod 400 illustrated in FIG. 4. Separately, in a polymer solutionpreparation step 615 of the method 600 illustrated in FIG. 6, a polymersolution is prepared as described herein, e.g. with respect to picklingstep 330 of the method 300 illustrated in FIG. 3. In a combining step620 of the method 600 illustrated in FIG. 6, the inactivated fungalbiomass is combined with the polymer solution as described herein, e.g.with respect to pickling step 330 of the method 300 illustrated in FIG.3 and/or pickling step 435 of the method 400 illustrated in FIG. 4. Inan initial drying step 630 of the method 600 illustrated in FIG. 6, theinactivated fungal biomass is dried as described herein, e.g. withrespect to drying step 360 of the method 300 illustrated in FIG. 3and/or drying step 495 of the method 400 illustrated in FIG. 4. In alaminating step 640 of the method 600 illustrated in FIG. 6, theinactivated fungal biomass is laminated together with one or more otherinactivated fungal biomasses and/or layers of non-fungal textilematerials, by any suitable method, to form a composite fungal sheet. Ina heat-pressing step 650 of the method 600 illustrated in FIG. 6, thecomposite fungal sheet is heat-pressed as described herein, e.g. withrespect to heat-pressing step 370 of the method 300 illustrated in FIG.3 and/or heat-pressing step 485 of the method 400 illustrated in FIG. 4.In a finishing step 660 of the method 600 illustrated in FIG. 6, one ormore traditional leather finishing waxes or oils (e.g. carnauba wax,candelilla wax) or nitrocellulose may be added to the composite fungalsheet, in any suitable amount and for any suitable time. In a finaldrying step 670 of the method 600 illustrated in FIG. 6, the compositefungal sheet is dried as described herein, e.g. with respect to dryingstep 360 of the method 300 illustrated in FIG. 3 and/or drying step 495of the method 400 illustrated in FIG. 4, to form the fungal textilematerial.

Generally, the methods illustrated in FIGS. 3-5 utilize a series ofchemical washes, which are conducted with agitation to increasediffusion of chemical species into the fungal structure and soften thefeel of the finished fungal textile material. The liming steps of thesemethods swell the matrix of the fungal structure and cleave certainfungal proteins, allowing for better diffusion of chemical species intothe fungus and exposing chemically active sites for reaction in laterprocess steps. Vegetable tannins can then be effective to form largehydrogen bond networks and thus crosslink the fungal structure,providing a strength, color, smell, and/or chemical stabilitycharacteristic of true leather. As in the methods illustrated in FIGS. 1and 2, a strengthening polymer (e.g. chitosan) and another non-tannincrosslinker (e.g. citric acid) can be employed; in addition to havingthe effects described above with respect to FIGS. 1 and 2, thestrengthening polymer and non-tannin crosslinker can form complexes withthe tannin crosslinker. Likewise, a plasticizer (e.g. glycerol) can alsobe incorporated into the methods.

It is to be expressly understood that any one or more filamentous fungimay suitably be used to form fungal textile materials of the presentinvention, including but not limited to one or more filamentous fungibelonging to a phylum selected from the group consisting of Ascomycotaand Basidiomycota; one or more filamentous fungi belonging to an orderselected from the group consisting of Ustilaginales, Russulales,Agaricales, Pezizales, and Hypocreales; one or more filamentous fungibelonging to a family selected from the group consisting ofUstilaginaceae, Hericiaceae, Polyporaceae, Grifolaceae, Lyophyllaceae,Strophariaceae, Lycoperdaceae, Agaricaceae, Pleurotaceae,Physalacriaceae, Omphalotaceae, Tuberaceae, Morchellaceae,Sparassidaceae, Nectriaceae, and Cordycipitaceae; one or morefilamentous fungi belonging to a genus selected from the groupconsisting of Agaricus, Calocybe, Calvatia, Cordyceps, Disciotis, Fomes,Fusarium, Ganoderma, Grifola, Hericulum, Hypholoma, Hypsizygus,Morchella, Pholiota, Pleurotus, Polyporous, Sparassis, Stropharia,Tuber, Ustilago; and/or one or more filamentous fungi belonging to aspecies selected from the group consisting of Ustilago esculenta,Hericulum erinaceus, Polyporous squamosus, Grifola fondosa, Hypsizygusmarmoreus, Hypsizygus ulmarius, Calocybe gambosa, Pholiota nameko,Calvatia gigantea, Agaricus bisporus, Stropharia rugosoannulata,Hypholoma lateritium, Pleurotus eryngii, Pleurotus ostreatus, Pleurotusostreatus var. columbinus, Tuber borchii, Morchella esculenta, Morchellaconica, Morchella importuna, Sparassis crispa, Fusarium venenatum, MK7ATCC Accession Deposit No. PTA-10698, Disciotis venosa, and Cordycepsmilitaris.

In the practice of the present invention, inactivated fungal biomass isallowed to soak in and/or is agitated with the polymer, plasticizer,and/or crosslinker solution for a time sufficient to allow the mat to bepenetrated by and/or saturated with the polymer, plasticizer, and/orcrosslinker, generally at least about one hour. After soaking in and/orbeing agitated with the solution, the wet mat is removed from thesolution (whereupon excess solution may be removed from one or moresurfaces of the mat).

Plasticizers suitable for use in the fungal textile materials of thepresent invention include but are not limited to glycerol and estersthereof, polyethylene glycol, citric acid, oleic acid, oleic acidpolyols (e.g. mannitol, sorbitol) and esters thereof, epoxidizedtriglyceride vegetable oils (e.g. from soybean oil), castor oil,pentaerythritol, fatty acid esters, carboxylic ester-based plasticizers,trimellitates, adipates, sebacates, maleates, biological plasticizers,and combinations and mixtures thereof. In the practice of the presentinvention, the plasticizer is typically present in the fungal textilematerial in an amount of between about 0.5 wt % and about 50 wt % or anysub-range between those values, including, by way of non-limitingexample, about 50 wt %, about 37.5 wt %, about 25 wt %, or about 12.5 wt%.

Polymers suitable for use in the fungal textile materials of the presentinvention include but are not limited to polyvinyl alcohol, chitosan,polyethylene glycol, polycaprolactones, polyacrylic acids, hyaluronicacid, alginates, and combinations and mixtures thereof In embodiments,two or more polymers may be included in any weight ratio between about99:1 and about 1:99; typically about 99:1, about 90:10, about 80:20,about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about20:80, about 10:90, or about 1:99; and more typically about 50:50. Aloading ratio of the textile composition may take any value betweenabout 99:1 and about 1:99; typically about 99:1, about 95:5, about90:10, about 85:15, about 80:20, about 75:25, about 70:30, about 65:35,about 60:40, about 55:45, about 50:50, about 45:55, about 40:60, about35:65, about 30:70, about 25:75, about 20:80, about 15:85, about 10:90,about 5:95, and about 1:99; and more typically about 70:30.

Crosslinkers suitable for use in the fungal textile materials of thepresent invention include but are not limited to citric acid, tannicacid, suberic acid, adipic acid, succinic acid, extracted vegetabletannins, glyoxal, and combinations and mixtures thereof. In embodiments,the fungal textile material may comprise proteins crosslinked withisopeptide bonds, the formation of which may in some embodiments becatalyzed by transglutaminase.

Relative amounts of filamentous fungus, plasticizer, polymer,crosslinker, additional components, etc. in the fungal textile materialsof the present invention may be selected to provide a fungal textilematerial having one or more desired physical, mechanical, sensory (e.g.olfactory, tactile, etc.) and/or aesthetic characteristics. Inembodiments, a scented additive, e.g. a leather fragrance oil, may beadded to the fungal textile material to provide a desired olfactorycharacteristic, e.g. a leather-like aroma, to the fungal textilematerial.

The filamentous fungus may make up about 20%-90%, or any sub-rangebetween those values, of the fungal textile material. In someembodiments, the filamentous fungus may make up between about 25-85%,about 30-80%, about 35-75% of the fungal textile material. For instance,in a non-limiting example, in some embodiments, it may make up betweenabout 40 wt % and about 60 wt % of the fungal textile material.

By way of further non-limiting examples, one or more polymers (e.g.chitosan) may make up between about 1 wt % and about 40 wt %, or anysub-range between those values, or between about 5 wt % and about 20 wt%, of the fungal textile material. By way of third non-limiting example,one or more crosslinkers (e.g. citric acid) may make up between about0.01 wt % and about 8 wt %, or any sub-range between those values, orbetween about 0.05 wt % and about 6 wt %, or between about 0.1 wt % andabout 4 wt % of the fungal textile material. By way of fourthnon-limiting example, one or more plasticizers (e.g. glycerol) may makeup between about 0.5 wt % and about 80 wt %, or any sub-range betweenthose values, or between about 9 wt % and about 60 wt %, or betweenabout 17.5 wt % and about 40 wt % of the fungal textile material.

Embodiments of the present invention include fungal textile materials,and particularly fungal leather analog materials, having engineeredand/or tuned thermal properties. By way of first non-limiting example,the thermal effusivity of the fungal textile material, i.e. the rate atwhich the fungal textile material exchanges heat with its surroundings,may be engineered or tuned according to the present invention. By way ofsecond non-limiting example, the thermal conductivity of the fungaltextile material, i.e. the quantity of heat transferred through thefungal textile material, may be engineered or tuned according to thepresent invention. By way of third non-limiting example, the heatcapacity, i.e. the amount of heat to be supplied to a given mass of thefungal textile material to produce a unit change in its temperature, maybe engineered or tuned according to the present invention. Thevolumetric heat capacity of the fungal textile material, i.e. thequantity of heat a volume of the fungal textile material can store, maybe engineered or tuned according to the present invention. The abilityto thermally engineer and/or tune the fungal textile materials allow thefungal textile material to have a desired “heat feel” and thusrepresents a major improvement over fungal or other non-animal textilematerials of the prior art, which frequently suffer from a drawback of“feeling cold” (i.e. having poor thermal properties) to a user and/oroffering insufficient insulation, e.g. to a wearer of an article ofclothing made from the fungal textile material; the present inventionthus allows for the creation of, e.g., fungal textiles that retain agreater quantity of heat and are thus suitable for use in articles ofwinter clothing. One further advantage and benefit of the presentinvention lies in the ability to produce textile materials that may havea combination of two or more of these or other thermal properties notachievable by conventional textile materials, e.g. it is possible toincrease one thermal property while increasing, holding constant, ordecreasing one or more other thermal properties, and/or it is possibleto hold one thermal property constant while increasing, holdingconstant, or decreasing one or more other thermal properties, and/or itis possible to decrease one thermal property while increasing, holdingconstant, or decreasing one or more other thermal properties.

Thermal properties of fungal textile materials of the present inventionmay be engineered or tuned by including in the fungal textile material athermal dopant. Thermal dopants suitable for use in the fungal textilematerials of the present invention include materials that modify one ormore of thermal effusivity, thermal conductivity, and heat capacity ofthe fungal textile material, as compared to the fungal textile materialin the absence of the thermal dopant. Such thermal dopants may comprise,but are not necessarily limited to, polymeric, ceramic, and metallicmaterials having known thermal properties, and/or any other materialhaving a desired thermally conductive and/or thermally insulativeproperty. Further non-limiting examples of thermal dopants suitable foruse in the present invention include activated charcoal, aluminum oxide,bentonite, diatomaceous earth, ethylene vinyl acetate, lignin,nanosilica, polycaprolactone, polylactic acid, silicone, and yttriumoxide. In some embodiments, the thermal dopant may comprise anengineered coating and/or an engineered spatial distribution ofthermally conductive and/or thermally insulative materials throughoutthe fungal textile material to produce a preselected thermal profile.

In the practice of the present invention, thermal dopants may be addedand/or introduced into the fungal textile material at any suitable pointin the manufacturing method. As a first non-limiting example, a thermaldopant may be provided in the polymer solution, i.e. combined with thepolymer and solvent before being subsequently combined with a fungalbiomass. As a second non-limiting example, a thermal dopant may becombined with the inactivated fungal biomass, water, and optionalpigment before or during a size-reducing and/or blending/homogenizingstep of the manufacturing method. As a third non-limiting example, athermal dopant may be added to the mixture of the fungal paste and thepolymer solution while the paste/polymer solution mixture is beingstirred and/or heated. As a fourth non-limiting example, a thermaldopant, and in some embodiments an engineered or designed spatialpattern or structure of a thermal dopant, may be integrated with thefungal textile material. As a fifth non-limiting example, a thermaldopant may be added before or during a casting step, e.g. by providingthe thermal dopant in a tray or mold in which the sheet is to be cast orby sprinkling or otherwise distributing particles of a dopant over asurface of the fungal material after casting. As a sixth non-limitingexample, a thermal dopant may be added to the fungal textile materialafter the fungal textile material has been cured.

The amount of the thermal dopant may be selected to provide a desiredthermal property to the resulting fungal textile material withoutcompromising other material properties (e.g. flexibility, tensilestrength, etc.) of the fungal textile material. Typically, thermaldopants, when provided, may make up between about 0.1 wt % and about25%, or any sub-range between those values, of the fungal textilematerial. In some embodiments, the dopants may be present at about 0.1to about 20 wt %, or at about 0.1 to about 15 wt % of the fungal textilematerial. For instance, in various embodiments, the dopants may make upabout 1 wt %, about 2 wt %, about 3 wt %, about 4 wt %, about 5 wt %,about 6 wt %, about 7 wt %, about 8 wt %, about 9 wt %, about 10 wt %,about 11 wt %, about 12 wt %, about 13 wt %, about 14 wt %, about 15 wt%, or any tenth of a weight percent between 0.1 and 25, of the fungaltextile material.

In embodiments, the fungal composition formed after mixing with thepolymer solution may be cast to at least partially overlie a scaffold orsubstrate comprising a thermal dopant. In embodiments, a force may beapplied to at least one of the fungal composition and the scaffold orsubstrate to provide a heterogeneous spatial distribution of the fungalcomposition and the scaffold or substrate in the cast sheet. Inembodiments, the fungal composition and a thermal dopant may each beselectively applied to predetermined regions of a casting area toprovide a heterogeneous spatial distribution of the blended compositionand the thermal dopant in the cast sheet. In embodiments, the cast sheetmay comprise a multilayer structure having at least a first layer and asecond layer, the first layer comprising at least a portion of thefungal composition and the second layer comprising at least a portion ofthe thermal dopant.

Embodiments of the present invention include articles of clothingpartially or completely constructed of a fungal textile material of theinvention. Such articles of clothing include, by way of non-limitingexample, protective garments, shirts, pants, shorts, jackets, coats,belts, hats, gloves, shoes, boots, sandals, flip-flops, watch straps,and aprons.

Embodiments of the present invention include accessory items partiallyor completely constructed of a fungal textile material of the invention.Such accessory items include, by way of non-limiting example, wallets,purses, cases, suitcases, luggage items, bags, backpacks, and hip packs.

Embodiments of the present invention include furniture items partiallyor completely constructed of a fungal textile material of the invention.Such furniture items include, by way of non-limiting example, chairs,recliners, couches, sofas, loveseats, and ottomans.

Embodiments of the present invention include coverings partially orcompletely constructed of a fungal textile material of the invention.Such coverings include, by way of non-limiting example, coverings forautomobile seats, airplane seats and train seats.

Fungal textile materials according to the present invention may bemanufactured such that they are characterized by a desired material,mechanical, and/or physical property. As a first non-limiting example,fungal textile materials may be manufactured to have a desired tensilestrength, which may in embodiments be at least about 15 MPa or betweenabout 4 M Pa and about 15 MPa., or any sub-range between those values.As a second non-limiting example, fungal textile materials may bemanufactured to have a desired strain at break, which may in embodimentsbe between about 50 percent and about 60 percent, or between about 10percent and about 70 percent, or any sub-range between those values. Asa third non-limiting example, fungal textile materials may bemanufactured to have a desired degree of swelling, which may inembodiments be between about 50 percent and about 60 percent, or betweenabout 30 percent and about 120 percent, or any sub-range between thosevalues. As a fourth non-limiting example, fungal textile materials maybe manufactured to have a desired mass loss upon soaking, which may inembodiments be no more than about 5 percent. As a fifth non-limitingexamples, fungal textile materials may be manufactured to have a desiredaverage fungal particle size, which may in embodiments be no more thanabout 25 nanometers, no more than about 50 nanometers, no more thanabout 75 nanometers, no more than about 100 nanometers, no more thanabout 125 nanometers, no more than about 150 nanometers, no more thanabout 175 nanometers, no more than about 200 nanometers, no more thanabout 225 nanometers, no more than about 250 nanometers, no more thanabout 275 nanometers, no more than about 300 nanometers, no more thanabout 325 nanometers, no more than about 350 nanometers, no more thanabout 375 nanometers, no more than about 400 nanometers, no more thanabout 425 nanometers, no more than about 2 micrometers, no more thanabout 4 micrometers, no more than about 6 micrometers, no more thanabout 8 micrometers, no more than about 10 micrometers, no more thanabout 15 micrometers, no more than about 20 micrometers, no more thanabout 30 micrometers, no more than about 40 micrometers, no more thanabout 50 micrometers, no more than about 75 micrometers, no more thanabout 100 micrometers, no more than about 150 micrometers, no more thanabout 200 micrometers, no more than about 250 micrometers, no more thanabout 300 nanometers, no more than about 400 micrometers, no more thanabout 500 micrometers and no more than about 750 micrometers. In someembodiments, the fungal biomass may comprise fungal filaments having alength of at least about 1 centimeter, at least about 2 centimeters, atleast about 3 centimeters, at least about 4 centimeters, at least about5 centimeters, at least about 6 centimeters, at least about 7centimeters, at least about 8 centimeters, at least about 9 centimeters,at least about 10 centimeters, at least about 20 centimeters, at leastabout 30 centimeters, at least about 40 centimeters, at least about 50centimeters, at least about 60 centimeters, at least about 70centimeters, at least about 80 centimeters, or at least about 90centimeters. As a sixth non-limiting example, fungal textile materialsmay be manufactured to have a desired type of particle sizedistribution, which may in embodiments be a bimodal, approximatelybimodal, trimodal, or approximately trimodal particle size distribution.As a seventh non-limiting example, fungal textile materials may bemanufactured to have a desired tear strength, which may in embodimentsbe between about 5 N/mm and about 25 N/mm, or any sub-range betweenthose values. As an eighth non-limiting example, fungal textilematerials may be manufactured to have a desired color fastness to wetrub, dry rub, and/or xenon light of at least about 4 in grey scale. As aninth non-limiting example, fungal textile materials may be manufacturedto have a desired flexural rigidity, which may in embodiments be no morethan about 5 gram-centimeters. One of the advantages and benefits of thepresent invention lies in the ability to produce textile materials thatmay have a combination of two or more of these or other material,mechanical, and/or physical properties not achievable by conventionaltextile materials, e.g. a combination of high tear strength (in someembodiments, at least about 1 N/mm, or at least about 2 N/mm, or atleast about 3 N/mm, or at least about 4 N/mm, or at least about 5 N/mm,or at least about 6 N/mm, or at least about 7 N/mm, or at least about 8N/mm, or at least about 9 N/mm, or at least about 10 N/mm, or at leastabout 11 N/mm, or at least about 12 N/mm, or at least about 13 N/mm, orat least about 14 N/mm, or at least about 15 N/mm, or at least about 16N/mm, or at least about 17 N/mm, or at least about 18 N/mm, or at leastabout 19 N/mm, or at least about 20 N/mm) and low flexural rigidity (insome embodiments, no more than about 10 gram-centimeters, or no morethan about 9 gram-centimeters, or no more than about 8 gram-centimeters,or no more than about 7 gram-centimeters, or no more than about 6gram-centimeters, or no more than about 5 gram-centimeters, or no morethan about 4 gram-centimeters, or no more than about 3 gram-centimeters,or no more than about 2 gram-centimeters, or no more than about 1gram-centimeter).

In some embodiments, a fungal leather analog material made from asize-reduced inactivated fungal biomass and being devoid of anynon-fungal textile backing may be provided.

Such fungal leather analog materials may have any one or more of thefollowing properties: a thickness of between about 1 and about 2 mm orbetween about 1.15 and about 1.6 mm, a tear strength of between about 6and about 12 N or between about 7.4 and about 10.5 N, a tensile strengthof between about 3 and about 10 N/mm² or between about 4.7 and about 8.1N/mm², a flexural rigidity of between about 1 and about 11 g. cm, awater spotting grey scale rating of 4 to 5, a light color fastness bluewool rating of at least 4, a rub color fastness grey scale rating whendry of 4 to 5, and a rub color fastness grey scale rating when dry of 4to 5. Such fungal leather analog materials can easily take on varioustextures or embossments.

In some embodiments, a fungal leather analog material made from asize-reduced inactivated fungal biomass and having a non-fungal textilebacking adhered on one side may be provided. Such fungal leather analogmaterials may have any one or more of the following properties: athickness of between about 1 and about 3 mm or between about 1.95 andabout 2.09 mm, a tear strength of between about 20 and about 50 N orbetween about 33 and about 37 N, a tensile strength of between about 3and about 10 N/mm² or between about 5.8 and about 6.8 N/mm², a flexuralrigidity of between about 1 and about 11 g·cm, a water spotting greyscale rating of 4 to 5, a light color fastness blue wool rating of atleast 4, a rub color fastness grey scale rating when dry of 4 to 5, anda rub color fastness grey scale rating when dry of 4 to 5. Such fungalleather analog materials can easily take on various textures orembossments.

In some embodiments, a composite fungal leather analog material madefrom a size-reduced inactivated fungal biomass and having a non-fungaltextile layer adhered between two layers of fungal material (i.e. amaterial in which the non-fungal layer is “sandwiched” between fungallayers) may be provided. Such fungal leather analog materials may haveany one or more of the following properties: a thickness of betweenabout 1 and about 4 mm or between about 2.2 and about 2.8 mm, a tearstrength of between about 25 and about 60 N or between about 34 andabout 52 N, a tensile strength of between about 7 and about 14 N/mm² orbetween about 8.7 and about 11.4 N/mm², a flexural rigidity of betweenabout 1 and about 11 g. cm, a water spotting grey scale rating of 4 to5, a light color fastness blue wool rating of at least 4, a rub colorfastness grey scale rating when dry of 4 to 5, and a rub color fastnessgrey scale rating when dry of 4 to 5. Such fungal leather analogmaterials can easily take on various textures or embossments.

In some embodiments, a fungal leather analog material made from aninactivated fungal biomass taking the form of one or more intact orwhole biomats (e.g. a biomass produced by surface fermentation and notsubjected to size reduction) and being devoid of any non-fungal textilebacking may be provided. Such fungal leather analog materials may haveany one or more of the following properties: a thickness of betweenabout 0.1 and about 1.5 mm per biomat or between about 0.5 and about 0.9mm per biomat, a tear strength of between about 1 and about 3 N perbiomat, a tensile strength of about 3 N/mm² per biomat, a flexuralrigidity of between about 1 and about 11 g·cm, and a water spotting greyscale rating of 4 to 5. Such fungal leather analog materials can haveadvantages such as warmth, drapability, softness, appearance, and smellthat closely mimic the same qualities of true leather.

In some embodiments, fungal leather analog materials made frominactivated fungal biomass, and methods of manufacture thereof, mayprovide environment advantages and benefits relative to true leather inaddition to the non-use of animal products. Particularly, the methods ofmanufacture of the present invention may generate no, or at leastsmaller quantities of, highly toxic or environmentally hazardousmaterials used in traditional leather tanning processes, such ashexavalent chromium compounds. Additionally, leather analog materialsaccording to the present invention may be biodegradable, i.e. biodegrademore quickly under a given set of conditions than true leather.

One feature of the invention is the ability to permit various chemicalcomponents (a polymer, a crosslinker, etc.) to infiltrate the mycelialmatrix of an inactivated fungal biomass. Where the inactivated fungalbiomass is a size-reduced fungal biomass, this infiltration may be theresult of the high surface area of fungal particles in contact with theinfiltrating fluid(s). Where the inactivated fungal biomass is an intactor cohesive fungal biomass (e.g. a biomat produced by surfacefermentation), this infiltration may be achieved by any one or more ofan extended time of contact between the fungal biomass and the fluid(s),agitation of the fungal biomass together with the fluid(s), applicationof sub- or superatmospheric pressure to the fungal biomass and thefluid(s), and so on.

It is one aspect of the present invention to provide a method forpreparing a durable sheet material comprising fungal biomass, comprising(a) combining an inactivated fungal biomass with at least one componentselected from the group consisting of a plasticizer, a polymer, acrosslinker, and a dye to form a combined composition; (b) casting thecombined composition to form a cast sheet; (c) removing solvent from thecast sheet; and (d) curing the cast sheet to form the durable sheetmaterial. It is to be expressly understood that this method can be usedin conjunction with either intact cohesive biomass (e.g. a biomatproduced by surface fermentation) or size-reduced fungal biomass.

In embodiments, step (d) may comprise drying the cast sheet.

In embodiments, step (d) may comprise initiating a chemical reactionwithin or on a surface of the cast sheet.

In embodiments, the method may further comprise adding at least one of anatural fiber material, a synthetic material, and combinations thereofto the blended composition. The natural fiber material may, but neednot, comprise a cellulosic material. The natural fiber material may, butneed not, comprise cotton fiber. The at least one of the natural fibermaterial and the synthetic material may, but need not, be in the form ofa plurality of particles, a sheet, or a combination thereof.

In embodiments, the method may further comprise inactivating a fungalbiomass to form the inactivated fungal biomass.

In embodiments, the method may further comprise size-reducing a fungalbiomass.

In embodiments, the method may further comprise adding a thermal dopantto at least one of the inactivated fungal biomass, the blendedcomposition, and the cast sheet.

It is another aspect of the present invention to provide a method forpreparing a durable sheet material comprising fungal biomass, comprising(a) contacting an inactivated fungal biomass with a solution comprisingat least one component selected from the group consisting of aplasticizer, a polymer, a crosslinker, and a dye; (b) removing a solventfrom the biomass; and (c) curing the biomass to form the durable sheetmaterial.

In embodiments, the method may further comprise inactivating a fungalbiomass to form the inactivated fungal biomass.

It is another aspect of the present invention to provide a textilecomposition, comprising an inactivated fungal biomass; and at least onecomponent selected from the group consisting of a plasticizer, apolymer, a crosslinker, and a dye.

In embodiments, the textile composition may comprise a plasticizer, apolymer, and a crosslinker.

In embodiments, the fungal biomass may comprise a fungus belonging to aphylum selected from the group consisting of Ascomycota andBasidiomycota.

In embodiments, the fungal biomass may comprise a fungus belonging to agenus selected from the group consisting of Fusarium, Fomes, andGanoderma. The fungus may, but need not, belong to a species selectedfrom the group consisting of Fusarium venenatum, Fomes fomentarius,Ganoderma applanatum, Ganoderma curtisii, Ganoderma formosanum,Ganoderma nei-japonicum, Ganoderma resinaceum, Ganoderma sinense, andGanoderma tsugae.

In embodiments, the fungal biomass may comprise a fungus selected fromthe group consisting of Fusarium venenatum and MK7 ATCC AccessionDeposit No. PTA-10698.

In embodiments, the plasticizer may comprise at least one selected fromthe group consisting of glycerol, polyethylene glycol, citric acid, andoleic acid. The plasticizer may, but need not, comprise glycerol. Theglycerol may, but need not, be present in the textile composition in anamount of between about 0.5 wt % and about 50 wt %. %, or any sub-rangebetween those values. In embodiments, the glycerol may, but need not, bepresent in an amount of about 50 wt %, about 37.5 wt %, about 25 wt %,or about 12.5 wt %.

In embodiments, the polymer may comprise at least one selected from thegroup consisting of polyvinyl alcohol, chitosan, polyethylene glycol,and hyaluronic acid. The polymer may, but need not, comprise polyvinylalcohol. The polymer may, but need not, comprise chitosan. The polymermay, but need not, comprise polyvinyl alcohol and chitosan. A weightratio of polyvinyl alcohol to chitosan may, but need not, be selectedfrom the group consisting of about 99:1, about 90:10, about 80:20, about70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80,about 10:90, and about 1:99 or any range formed by two of those ratios.The weight ratio of polyvinyl alcohol to chitosan may, but need not, beabout 50:50.

In embodiments, the textile composition may comprise a polymer, whereina loading ratio of the textile composition is selected from the groupconsisting of about 99:1, about 95:5, about 90:10, about 85:15, about80:20, about 75:25, about 70:30, about 65:35, about 60:40, about 55:45,about 50:50, about 45:55, about 40:60, about 35:65, about 30:70, about25:75, about 20:80, about 15:85, about 10:90, about 5:95, and about 1:99or any range formed by two of those ratios. The loading ratio may, butneed not, be about 70:30.

In embodiments, the crosslinker may comprise at least one selected fromthe group consisting of citric acid, tannic acid, suberic acid, adipicacid, succinic acid, glyoxal, and extracted vegetable tannins. Thecrosslinker may, but need not, comprise adipic acid.

It is another aspect of the present invention to provide an article ofclothing, comprising a textile composition of the invention.

In embodiments, the article may be a protective garment.

In embodiments, the article of clothing may be selected from the groupconsisting of a shirt, a pant, a short, a jacket, a coat, a belt, a hat,a glove, a shoe, a boot, a sandal, a flip-flop, a watch strap, and anapron.

It is another aspect of the present invention to provide an accessoryitem, comprising a textile composition of the invention.

In embodiments, the accessory item may be selected from the groupconsisting of a wallet, a purse, a case, a suitcase, a luggage item, abag, a backpack, and a hip pack.

It is another aspect of the present invention to provide a furnitureitem, comprising a textile composition of the present invention.

In embodiments, the furniture item may be selected from the groupconsisting of a chair, a recliner, a couch, a sofa, a loveseat, anottoman, and a vehicle seat.

In embodiments, the textile composition may have a tensile strength ofat least about 15 MPa.

In embodiments, the textile composition may have a strain at break ofbetween about 30 percent and about 60 percent.

In embodiments, the textile composition may have a degree of swelling ofbetween about 30 percent and about 60 percent.

In embodiments, the textile composition may have a mass loss uponsoaking of no more than about 30 percent.

In embodiments, the fungal biomass may have an average particle sizeselected from the group consisting of no more than about 25 nanometers,no more than about 50 nanometers, no more than about 75 nanometers, nomore than about 100 nanometers, no more than about 125 nanometers, nomore than about 150 nanometers, no more than about 175 nanometers, nomore than about 200 nanometers, no more than about 225 nanometers, nomore than about 250 nanometers, no more than about 275 nanometers, nomore than about 300 nanometers, no more than about 325 nanometers, nomore than about 350 nanometers, no more than about 375 nanometers, nomore than about 400 nanometers, no more than about 425 nanometers, nomore than about 2 micrometers, no more than about 4 micrometers, no morethan about 6 micrometers, no more than about 8 micrometers, no more thanabout 10 micrometers, no more than about 15 micrometers, no more thanabout 20 micrometers, no more than about 30 micrometers, no more thanabout 40 micrometers, no more than about 50 micrometers, no more thanabout 75 micrometers, no more than about 100 micrometers, no more thanabout 150 micrometers, no more than about 200 micrometers, no more thanabout 250 micrometers, no more than about 300 nanometers, no more thanabout 400 micrometers, no more than about 500 micrometers and no morethan about 750 micrometers.

In embodiments, the fungal biomass may have a bimodal or approximatelybimodal particle size distribution.

In embodiments, the fungal biomass may have a trimodal or approximatelytrimodal particle size distribution.

In embodiments, the textile composition may comprise transglutaminase.

In embodiments, the textile composition may comprise proteinscrosslinked with isopeptide bonds. Formation of the crosslinkingisopeptide bonds may, but need not, be catalyzed by transglutaminase.

In embodiments, the textile composition may further comprise a thermaldopant. The thermal dopant may, but need not, be selected from the groupconsisting of activated charcoal, aluminum oxide, bentonite,diatomaceous earth, lignin, nanosilica, polycaprolactone, polylacticacid, silicone, and yttrium oxide.

It is another aspect of the present invention to provide a method forpreparing a durable sheet material comprising fungal biomass, comprising(a) homogenizing an inactivated fungal biomass with a fluid comprisingwater to form a fungal paste; (b) combining the fungal paste with anaqueous solution comprising a polymer to form a blended composition; (c)casting the blended composition to form a cast sheet; (d) removingsolvent from the cast sheet; and (e) curing the cast sheet to form thedurable sheet material. It is to be expressly understood that fungalbiomass suitable for use in this method may be produced by any of anumber of methods known in the art and disclosed herein, including butnot limited to surface fermentation methods, submerged fermentationmethods, solid-substrate submerged fermentation (SSSF) methods, andmethods as disclosed in the '474 publication.

In embodiments, the fluid of step (a) may further comprise a pigment.

In embodiments, the method may further comprise inactivating a fungalbiomass to provide the inactivated fungal biomass.

In embodiments, step (a) may further comprise simultaneouslysize-reducing the inactivated fungal biomass.

In embodiments, the inactivated fungal biomass may be a size-reducedfungal biomass.

In embodiments, the polymer may comprise chitosan.

In embodiments, the aqueous solution of step (b) may further comprise atleast one of a crosslinker, a plasticizer, a solubilizer, and a pHadjusting agent. The aqueous solution may, but need not, comprise acrosslinker, wherein the crosslinker comprises citric acid. The aqueoussolution may, but need not, comprise a plasticizer, wherein theplasticizer comprises glycerol.

In embodiments, the method may further comprise, between steps (a) and(b), degassing the fungal paste.

In embodiments, the method may further comprise, between steps (b) and(c), degassing the blended composition.

In embodiments, the method may further comprise adding at least onethermal dopant to at least one of the inactivated fungal biomass, thefluid of step (a), the fungal paste, the aqueous solution of step (b),the blended composition, the cast sheet, and a tray, mold, or othervessel into which the blended composition is cast in step (c).

In embodiments of any of the above methods, the fungal biomass may beproduced by a method comprising culturing a fungal inoculum by at leastone of surface fermentation, submerged fermentation, solid-substratesubmerged fermentation, and a fermentation method as described in the'474 publication. The fungal biomass may, but need not, be a biomat.

Embodiments of any of the above methods may further comprise maintainingor introducing at least one bubble of a gas.

Embodiments of the above textile compositions may comprise at least onebubble of a gas.

In embodiments, the fungal biomass may comprise fungal filaments havinga length of at least about 1 centimeter, at least about 2 centimeters,at least about 3 centimeters, at least about 4 centimeters, at leastabout 5 centimeters, at least about 6 centimeters, at least about 7centimeters, at least about 8 centimeters, at least about 9 centimeters,at least about 10 centimeters, at least about 20 centimeters, at leastabout 30 centimeters, at least about 40 centimeters, at least about 50centimeters, at least about 60 centimeters, at least about 70centimeters, at least about 80 centimeters, at least about 90centimeters, at least about 100 centimeters, at least about 200centimeters, at least about 300 centimeters, at least about 400centimeters, at least about 500 centimeters, at least about 600centimeters, at least about 700 centimeters, at least about 800centimeters, or at least about 900 centimeters.

In embodiments, the fungal biomass may comprise fungal filaments havinga length of no more than about 1 centimeter, no more than about 9millimeters, no more than about 8 about millimeters, no more than about7 millimeters, no more than about 6 millimeters, no more than about 5millimeters, no more than about 4 millimeters, no more than about 3millimeters, no more than about 2 millimeters, no more than about 1millimeter, no more than about 900 micrometers, no more than about 800micrometers, no more than about 700 micrometers, no more than about 600micrometers, no more than about 500 micrometers, no more than about 400micrometers, no more than about 300 micrometers, no more than about 200micrometers, no more than about 100 micrometers, no more than about 90micrometers, no more than about 80 micrometers, no more than about 70micrometers, no more than about 60 micrometers, no more than about 50micrometers, no more than about 40 micrometers, no more than about 30micrometers, no more than about 20 micrometers, no more than about 10micrometers, no more than about 9 micrometers, no more than about 8micrometers, no more than about 7 micrometers, no more than about 6micrometers, no more than about 5 micrometers, no more than about 4micrometers, no more than about 3 micrometers, no more than about 2micrometers, or no more than about 1 micrometer.

The invention is further illustratively described by way of thefollowing non-limiting Examples.

EXAMPLE 1 Textile Material Manufacturing Process

Fungal textile materials according to the present invention may, inembodiments, be made according to the methods described in this Example.In particular, the methods described in this Example may be employed tomanufacture a leather analog textile material, i.e. a fungal textilematerial that may replicate, simulate, and/or substitute for trueleather.

The first step or steps in these methods of making a fungal textilematerial generally include obtaining a mat of fungal material,comprising fungal mycelia, from a suitable reactor, which may inembodiments entail producing a fungal biomat according to the methodsdescribed in the '050 application, the '626 application, the '421application, and/or the '474 publication. These mats are theninactivated, in some embodiments by steaming for not less than 30minutes, and the inactivated mat may then be cut into a desired size andgeometry. In some embodiments, the mat may be partially or completelydried in a dehydrator at elevated temperature, e.g. between about 130°F. and about 160° F.

The inactivated mat is then placed into a solution of one or morecomponents selected to impart a desired characteristic to the finalfungal textile material. Generally, the solution comprises one or moreof a polymer, a plasticizer, and a crosslinker. Polymers suitable foruse in solution according to the present invention include but are notlimited to polyvinyl alcohol, chitosan, polyethylene glycol, hyaluronicacid, polycaprolactones, polyacrylic acids, and combinations andmixtures thereof. Plasticizers suitable for use in solution according tothe present invention include but are not limited to glycerol and estersthereof, polyethylene glycol, citric acid, oleic acid, oleic acidpolyols (e.g. mannitol, sorbitol) and esters thereof, epoxidizedtriglyceride vegetable oils (e.g. from soybean oil), castor oil,pentaerythritol, fatty acid esters, carboxylic ester-based plasticizers,trimellitates, adipates, sebacates, maleates, biological plasticizers,and combinations and mixtures thereof. Crosslinkers suitable for use insolution according to the present invention include but are not limitedto citric acid, tannic acid, suberic acid, adipic acid, succinic acid,extracted vegetable tannins, glyoxal, and combinations and mixturesthereof.

The inactivated mat is allowed to soak in the polymer, plasticizer,and/or crosslinker solution for a time sufficient to allow the mat to bepenetrated by and/or saturated with the polymer, plasticizer, and/orcrosslinker, generally at least about two hours and most typically about24 hours. After soaking in the solution, the wet mat is removed from thesolution (whereupon excess solution may be removed from one or moresurfaces of the mat).

An optional step in the methods of the present invention, which may bepreferable in some embodiments, includes lamination of two or more matsafter soaking. In the practice of the present invention, mats may belaminated by vertically stacking two or more mats or arranging the matsin any desired spatial orientation (horizontal vs. vertical, parallelvs. perpendicular vs. oblique, etc.), which may in some cases includenatural fibers in addition to the fungal mycelia, and soaking thevertically stacked mats in a polymer solution, which may be the same asor different from the solution used for the earlier soaking step.Generally, lamination of two or more mats according to the presentinvention includes the removal of air bubbles trapped between layers,e.g. by pressing the stacked mats, by rolling, by vacuum extraction,etc.

Wet mats (or laminates of mats) are then dried, generally for betweenabout 30 minutes and about 120 minutes, in a dehydrator at elevatedtemperature, e.g. between about 130° F. and about 160° F., to removesubstantially all of the liquid from an outer surface of the mat (orlaminate) but retain at least some liquid in an interior of the mat (orlaminate). The mats are then removed from the dehydrator and, in someembodiments, heat-pressed, e.g. between textured silicon molds, atelevated temperature (e.g. about 130° C.); typically, mats areheat-pressed in intervals of between about 20 seconds and about 30seconds for a total time of between about 3 minutes and about 10minutes.

EXAMPLE 2 Fungal Growth Through Fibers

Fungal textile materials according to the present invention may, inembodiments, be made according to the methods described in this Example.In particular, the methods described in this Example may be employed tomanufacture a textile material that incorporates both filamentous fungusand other natural or synthetic fibers.

The first step or steps in these methods of making a fungal textilematerial generally include providing a growth medium for filamentousfungus, which may in embodiments include growth media as described inthe '050 application, the '626 application, the '421 application, and/orthe '474 publication, but which may also include other types of growthmedium. Particularly, a growth medium may be formulated with analternative carbon source or different carbon content, which may inembodiments promote the consumption of natural fibers by the fungus tobe cultured in the growth medium. By way of non-limiting example,conventional growth media may be altered by replacing glycerol withhydrolyzed cellulose, crystalline cellulose, or other cellulosiccompounds to promote production of cellulase enzymes by the filamentousfungus. By way of further non-limiting example, the total amount ofcellulosic material may be carefully controlled, e.g. to about 10 wt %of the growth medium, to provide for desired growth characteristics ofthe filamentous fungus. After the growth medium is prepared, it isgenerally boiled for a period of no less than 30 minutes to eliminatecompetitive or pathogenic microorganisms, then sealed and left to cool.The cooled medium is typically pH adjusted, e.g. using hydrogenchloride, and inoculated with an inoculum of a filamentous fungus (e.g.MK7 ATCC Accession Deposit No. PTA-10698) at a rate of about 5 vol %;the medium is generally stirred to provide uniform dispersal of thefungal inoculum.

A reactor for the production of filamentous fungus biomass is preparedby providing a sanitary reactor, e.g. a Saran wrap reactor or the like,and cleaning and/or sterilizing an interior (e.g. walls, doors, racks,trays, etc.) of the reactor (e.g. with ethanol). Separately, naturalfibers which are to serve as a substrate and/or structural material forthe fungal textile material are placed into one or more Pyrex trays,generally at a rate of about 0.5 grams to about 5 grams per tray,covered in aluminum foil, and dry-autoclaved to eliminate competitive orpathogenic microorganisms, then allowed to cool; the Pyrex trays arethen placed into the cleaned reactor (generally onto trays of thereactor).

The inoculated medium is then poured or otherwise introduced into thePyrex trays in the reactor, generally at a rate of about 200 mL pertray. It is generally desirable to introduce the inoculated medium intoa corner of the Pyrex tray rather than its center, to allow the growthmedium to flow beneath the fibers within the Pyrex tray and thus toallow the fibers to float on the surface of the liquid medium. After anincubation period, generally between about three days and about threeweeks, each Pyrex tray contains a fungal biomass grown through thenatural substrate and/or structural fibers, which may then be harvestedfor further processing.

EXAMPLE 3 Incorporation of Oil(s)

In the production of true (i.e. non-fungal) leather, the leathermaterial is generally subjected to an oiling process, whereby theleather material is coated with one or more oils, or more commonly witha mixture of oil(s), an emulsifier, and a penetrating aid. This oilingprocess lubricates the leather and improves its ability to flex withoutcracking (dry leather fibers generally crack or break easily) and mayalso impart color and water resistance to the leather material. In thepractice of the present invention, oils may be likewise incorporatedinto fungal leather analog materials, or produced in situ by thefilamentous fungus itself during a fermentation process, to providesimilar advantages and benefits. This Example describes embodiments ofsuch oil incorporation processes for fungal leather analog materials.

In “emulsion” oil incorporation methods according to the presentinvention, one or more oils, fats, and/or waxes are provided. The oils,fats, and/or waxes may be selected for their utility as emulsifiersand/or surfactants (e.g. salts, soaps, and other amphiphilic molecules),and may include, by way of non-limiting example, any one or more ofsulfated castor oil, beeswax, coconut oil, vegetable oil, olive oil,linseed oil and oleic acid sulfated fish oil, sulfated canola oil,soybean oil, palm oil, fatty acids,. Emulsions formed with the aid ofsurfactants may provide more stable conditions for penetration of theleather; those of ordinary skill in the art can select an anionic,cationic, or non-ionic surfactant to improve the wetting action of theemulsion on the fibers of the leather material. These oils, fats, and/orwaxes are rapidly stirred in a vessel (e.g. via magnetic stir bar), andin some embodiments heat may be applied to melt one or more of the oils,fats, and/or waxes to ensure complete mixing, while water (preferablydeionized water) is gradually added to the mixture until a milkyemulsion is formed; most typically, water makes up between about 50 vol% and about 70 vol % of this emulsion. The stirring rate is subsequentlyreduced (e.g. via magnetic stir bar or orbital shaker), whereupon fungalleather analog materials according to the present invention areintroduced to the vessel. The fungal leather analog materials aregenerally allowed to remain in the agitated emulsion for a period ofbetween about 20 minutes and about four hours, then removed from theemulsion and allowed to air-dry for between about 24 hours and about 48hours. This oiling process may be carried out before, after, and/or inlieu of heat-pressing of the fungal leather analog material.

In “stuffing” oil incorporation methods according to the presentinvention, one or more liquefied oils or waxes, including but notlimited to oils or waxes suitable for use in the “emulsion” methoddescribed above, may be mechanically rubbed onto the surface of a fungalleather analog material to “work” the oils or waxes into the structureof the fungal leather analog material. As in the “emulsion” method, thefungal leather analog material is then allowed to air-dry for betweenabout 24 hours and about 48 hours, and the “stuffing” oiling process maybe carried out before, after, and/or in lieu of heat-pressing of thefungal leather analog material.

EXAMPLE 4 Vegetable Tanning

In the practice of the present invention, the use of a dicarboxylic acidas the crosslinker generally necessitates heat-pressing of the fungaltextile material because crosslinking of carboxylic acids to thechemical moieties found in the fungal textile material generally occursonly at elevated temperature (e.g. about 130° C.). As an alternative,natural tannins, such as tannins extracted from vegetable material orother plant material, may bond to and/or induce chemical bonding in(i.e. crosslink) the fungal textile material at lower temperature thandicarboxylic acids and thus eliminate the need for heat-pressing, whichmay improve the water resistance of the fungal textile material. Withoutwishing to be bound by any particular theory, it is believed thattannins interact with fungal textile materials in much the same way thatthey interact with animal hides or skins, i.e. bonding with proteinmoieties to improve the strength and degradation resistance of thematerial.

Elimination of the need to heat-press the fungal textile material mayhave further advantages and benefits for downstream processing. By wayof non-limiting example, oil incorporation processes (such as thosedescribed in Example 3) typically require a relatively “open” structureof the fungal textile material; heat-pressing closes the structure ofthe fungal textile material and thus makes it difficult for the oil topenetrate into the leather structure, and while the oil incorporationprocess may be performed before heat-pressing, this can in some casesinterfere with crosslinking reactions and/or cause oils to leach fromthe fungal textile material during heat-pressing. This Example describesembodiments of a process for crosslinking a fungal textile materialusing vegetable tannins to avoid these and other drawbacks.

In vegetable tanning methods according to the present invention, mats offungal biomass are produced by any suitable method, including but notlimited to methods as disclosed herein and/or in the '050 application,the '626 application, and/or the '421 application, and steamed asdescribed in Example 1. The steamed mats are washed one or more timeswith deionized water, brine, or a combination or mixture thereof, andthe washed mats are then placed in a solution containing tannincompounds. The tannin compounds may comprise any one or more commercialplant-extracted tannins and/or pure tannic acid, and generally make upbetween about 0.5 wt % and about 20 wt % of the tanning solution. Thefungal mats are generally allowed to remain in the tanning solution forbetween about one day and about 30 days, and in some embodiments thefungal mats may be transferred between two or more tanning solutions,e.g. tanning solutions having different compositions and/orconcentrations of tannin compounds, during the tanning process.

After tanning, the fungal mats may be oiled by any suitable method, e.g.one or both of the methods described in Example 3, and/or may besubjected to a plasticizing solution or process (e.g. using polyethyleneglycol (PEG) and/or glycerol as a plasticizer). The plasticized and/oroiled material is finally allowed to air-dry, generally for betweenabout 24 hours and about 72 hours. It is to be expressly understood thatfurther crosslinking, e.g. using dicarboxylic acids as crosslinkers,may, in embodiments, be performed after the vegetable tanning processdescribed in this Example.

EXAMPLE 5 Effects of Polymer-Plasticizer Ratio on Textile MaterialProperties

This Example describes the effect of a ratio of polymer to plasticizerin the solution of the present invention on material properties offungal textile materials, and fungal leather analog materialsparticularly. Polymers (i.e. long-chain molecules chemically bonded tobiological structures within the fungal textile material) improve thetensile strength of the fungal textile material, whereas plasticizers(i.e. smaller molecules that do not chemically bond to the biologicalstructures or the polymers) improve the flexibility and decrease thebrittleness of the fungal textile material. Thus, without wishing to bebound by any particular theory, it is believed that varying apolymer-to-plasticizer ratio (hereinafter “PP ratio”) may enable thoseof ordinary skill in the art to precisely control, select, or tunephysical properties of fungal textile materials produced according tothe present invention.

Biomats of MK7 ATCC Accession Deposit No. PTA-10698 (hereinafter “MK7”)were grown and steamed or boiled for 30 minutes to inactivate thefungus. The inactivated biomats were cut into approximately 4cm x 6cmrectangles, each of which was placed into a solution comprising both apolymer (either polyvinyl alcohol (PVA) or chitosan) and a plasticizer(glycerol) and left to soak overnight. After soaking, each rectangle wasdried for between about 45 minutes and about one hour in a tabletopdehydrator, then heat-pressed at 275° F. in 30-second intervals for atotal of four minutes. The samples were then air-dried at roomtemperature overnight and subsequently tested for degree of swelling(DOS), mass loss upon soaking (ML), tensile strength (TS), andsubjective flexibility (six evaluations, 0-10 scale). The results arepresented in Table 1.

TABLE 1 Material Properties of MK7 Leather Analog Samples with VaryingPolymer-Plasticizer Ratios Sample PVA Chitosan Glycerol PP DOS ML TS No.wt % wt % wt % ratio % % (MPa) Flexibility  1 10 0 15 0.67 113.84  39.933.88 9.00  2 10 0 10 1.00 127.91  32.23 5.09 8.83  3 10 0  8 1.25125.87  28.43 4.99 6.00  4 10 0  4 2.50 142.46  18.66 NA 1.67  5  5 0 100.50 120.84  33.21 3.29 8.83  6  5 0  8 0.63 130.81  27.92 3.89 8.67  7 5 0  4 1.25 125.05  21.89 NA 2.83  8  0 4 15 0.27 147.65  36.01 2.705.00  9  0 4 10 0.40 177.02  26.80 8.61 3.83 10  0 4  6 0.67 189.90 21.46 5.78 3.33 11  0 4  4 1.00 229.21  36.46 7.68 2.67 12  0 4  2 2.00164.35  17.39 NA 0.67 13  0 2 15 0.13 97.39 40.63 3.25 9.83 14  0 2 100.20 107.74  25.65 4.43 6.00 15  0 2  6 0.33 114.43  22.28 8.46 4.17 16 0 2  4 0.50 128.01  18.56 NA 0.67

Certain trends were evident regardless of the type of polymer (PVA vs.chitosan) used: an increase in tensile strength as the PP ratioincreases, an increase in the degree of swelling as the PP ratioincreases, a decrease in mass loss as the PP ratio increases, and adecrease in flexibility as the PP ratio increases. The introduction ofpolymers into MK7 biomats and subsequent heat-pressing causes theformation of covalent and non-covalent bonds between the fungal myceliaand polymer molecules. These polymer molecules also bind to one anotherto create an entanglement of bound structures. Plasticizing agents, suchas glycerol, are “free floating” molecules that remain unbound to boththe polymers and the MK7 structures and serve to block the formation ofchemical bonds between polymers and biomass. When plasticizers aresparsely present, more chemical bonding can occur, leading to materialsof increased strength and brittleness. When plasticizers are present inabundance, they block the formation of chemical bonds and lead tomaterials that are flexible but lack strength. This phenomenon isevidenced by the wide range of tensile strengths (2.70 MPa to 8.61 MPa)and the wide range of flexibilities (0.67 to 9.83 on a subjective 0-10scale) obtained by varying concentration of polymer to plasticizer.

Samples that utilized PVA as the polymer displayed more consistentresults in the mid-range of each testing parameter, while samples thatcontained chitosan as the polymer displayed less consistent results thatmore broadly spanned the extremes of the parameter ranges. This resultmay be partially attributable to the differences between steamed andboiled biomat samples; boiled samples were able to more uniformlyincorporate the contents of polymer solutions and therefore showedbetter performance, whereas steamed samples tended to be much morebrittle. It appeared that PVA was more readily and uniformly absorbed bysteamed biomats than chitosan, which may explain the more consistentdata obtained for PVA samples.

EXAMPLE 6 Effect of Glycerol Content on Textile Material Properties

The procedure of Example 5 was repeated, except that thepolymer/plasticizer solution contained no polymer (i.e. no PVA orchitosan) and the plasticizer (i.e. glycerol) content was varied toassess the effect of glycerol content on the material properties of thefungal textile material.

Over the range of glycerol concentration tested for MK7 leather samples,distinct trends in TS, strain at break (SAB), DOS, and ML were observed.The TS of MK7 leather, as illustrated in FIG. 7, was observed todecrease with an increase in glycerol concentration; a maximum TS of8.65 MPa was achieved for samples made from pre-boiled biomass and noadded glycerol, and a minimum TS value of 1.55 MPa was recorded for theraw biomass sample with an added glycerol concentration of 37.5%. TheSAB of MK7 leather, as illustrated in FIG. 8, was observed to increasewith an increase in glycerol concentration, although the samples madefrom pre-boiled biomass were less representative of this trend, mostlikely due to incomplete drying of some samples prior to strain testing.Additionally, the DOS of MK7 leather, as illustrated in FIG. 9, wasobserved to decrease with an increase in glycerol concentration, whilethe ML, as illustrated in FIG. 10, was observed to increase with anincrease in glycerol concentration.

Glycerol acts by disrupting polymer-polymer interactions, increasingfree space, and thus increasing the mobility of the polymer molecules.In MK7 leather embodiments according to the present invention, there isa mixture of PVA and/or chitosan polymers, along with native MK7 cellsand excreted biopolymers (EPS). In the absence of glycerol, the addedpolymers, cells, and biopolymers can form more hydrogen, ionic, andcovalent bonds to one another; molecular mobility and free space arelow, while bond concentration is high. In this state, the material ismore rigid and requires more energy to stretch or bend. Measured TS aretherefore higher, and strains lower, when the glycerol concentration islow, and vice versa.

EXAMPLE 7 Effect of Loading Ratio on Textile Material Properties

The procedure of Example 5 was repeated, except that thepolymer/plasticizer solution contained no plasticizer (i.e. no addedglycerol) and the total polymer content (i.e. total amount of PVA and/orchitosan) was varied to assess the effect of loading ratio on thematerial properties of the fungal textile material.

The TS of MK7 leather, as illustrated in FIG. 11, was shown to increasewith an increase in polymer concentration; in other words, at the lowestloading ratio, the highest tensile strength was observed and vice versa.The TS was observed to increase linearly with polymer concentration upto a polymer concentration of about 36.5%, and then a drop in TS wasobserved at a polymer concentration of about 47.5%. At polymerconcentrations of more than about 47.5%, the TS increased linearly to amaximum value of 6.89 MPa at a polymer concentration of 73%. Withoutwishing to be bound by any particular theory, it is believed that thiseffect is attributable to the many hydroxyl and amine groups present inPVA and chitosan molecules; these groups can form covalent andnon-covalent bonds with biological structures and with other polymermolecules. As the polymer concentration increases, so too does theconcentration of intermolecular bonds. High bond concentration thenleads to improved strength of material. Additionally, the unprocessedbiomass used to create the leather samples contains both remainingglycerol from media and EPS molecules created by the organism. Theglycerol, and likely some components of the EPS, serve as plasticizingagents to the leather structure. Based on the results of the glycerolconcentration experiment, it can be reasonably inferred that increasingthe biomass concentration, and thus the plasticizer concentration,likely causes a decrease in the TS of samples.

The SAB of MK7 leather samples, as illustrated in FIG. 12, was observedto increase linearly with polymer concentration to a maximum value of182% at a polymer concentration of 36.5%. As polymer concentrationincreased past 36.5%, the SAB was observed to then decrease linearly.Without wishing to be bound by any particular theory, it is believedthat this effect is attributable to the competing effects ofintermolecular bonding and plasticization within the leather structure.At high loading ratios, biomass and plasticizer concentrations are highwhile bond concentration is low, and the lack of intermolecular bondsleads to a material that has a low tension limit; thus, during tensiletesting, the tension limit is likely reached prior to significantmaterial strain, causing material failure at low strains. At the medianloading ratio (a polymer concentration of 37.5%), by contrast,significant intermolecular bonding likely occurs. Additionally, due tosignificant incorporation of biomass at the median loading ratio, thesamples are also significantly plasticized. These properties lead to amaterial with both a moderately high tension limit, and a moderatelyhigh strain limit. During tensile tests, the material can stretchsignificantly prior to reaching either its tension or strain limits. Atlower loading ratios, the samples are no longer significantlyplasticized. They contain a large concentration of polymers that formintermolecular bonds and therefore have a high tensile limit. However,the lack of plasticizing molecules leads to low strain limits andmaximum TS values are reached at lower corresponding SAB values.

A similar trend to that of SAB was observed for the DOS of leathersamples, as illustrated in FIG. 13. The DOS of samples increasedlinearly with an increase in polymer concentration to a maximum value of405% at a polymer concentration of 47.5%. As the polymer concentrationincreased past 47.5%, a linear decrease in DOS was observed. PVA andchitosan are known to form hydrogels, i.e. materials that comprise athree-dimensional mesh or network of physically and chemically boundpolymer molecules. When not fully crosslinked, the hydrogel network isflexible and contains spaces between polymer strands, and the hydrogelcan thus stretch and hold large amounts of water in the spaces betweenthe polymer strands. When fully crosslinked, spaces between polymerstrands are bound and the material is less able to flex as water isabsorbed. In this crosslinked state, hydrogels have less water holdingcapacity. Without wishing to be bound by any particular theory, it isbelieved that, at high loading ratios, there are fewer available bondingsites for water molecules due to the small quantity of polymermolecules. Therefore, at high loading ratios, there is less capacity toabsorb water. The maximum DOS values were observed at median polymerconcentrations; at these concentrations, there was a relatively highpolymer concentration, and a relatively high biomass concentration. Thebiomass contains absorbed glycerol and leads to a plasticized polymernetwork with low crosslinking and a high water holding capacity. As thepolymer concentration increases further, the plasticization effectdecreases with decreasing absorbed glycerol. This leads to a more highlycrosslinked material that is unable to hold as much water.

The ML values of MK7 leather however did not display any local maxima.Instead, the ML values, as illustrated in FIG. 14, were observed todecrease linearly with an increase in polymer concentration. Withoutwishing to be bound by any particular theory, it is believed that thiseffect is attributable to the decreasing amount of biomass associatedwith decreasing loading ratio. The biomass contains a large proportionof water-soluble compounds, which may diffuse into the aqueous phasewhen the biomass is soaked. The difference in mass from before and aftersoaking is therefore much higher for high loading ratio samples thatcontain significant masses of soluble compounds.

EXAMPLE 8 Effect of Polyvinyl Alcohol-Chitosan Ratio on Textile MaterialProperties

The procedure of Example 5 was repeated, except that thepolymer/plasticizer solution contained no plasticizer (i.e. no addedglycerol), the total polymer content (i.e. total amount of PVA and/orchitosan) was held constant, and the ratio of PVA to chitosan was variedto assess the effect of varying polymer compositions on the materialproperties of the fungal textile material.

The TS of MK7 leather samples, as illustrated in FIG. 15, were observedto have local maxima at PVA:chitosan weight ratios of 0:100, 50:50, and100:0. These points correspond to PVA concentrations of 0%, 11.7%, and23.4% and TS values at these points were 3.55 MPa, 3.53 MPa, and 4.32MPa, respectively. The SAB of samples, as illustrated in FIG. 16, wasobserved to have local maxima at the same PVA:chitosan ratios observedfor the TS. Values for SAB at these points were 143%, 138%, and 132%,respectively. Without wishing to be bound by any particular theory, itis possible that local TS maxima are observed when one polymer is absentand when the polymers are present in equal amounts because chemicalbonding of one polymer may be disrupted by the inclusion of smallamounts of the other polymer, i.e. chitosan, present in small amounts,may form aggregates within a larger matrix of PVA and vice versa. Thepolymer aggregates would compete with the biomass for binding sites ofthe large polymer matrix leading to decreased strength while alsodisrupting the ability of polymer molecules to move and stretch. Thiswould explain the low values of TS and SAB observed at PVA:chitosanratios of 80:20, 60:40, 40:60, and 20:80. When approximately equalamounts of each polymer are added, aggregates may not form and ahomogenous polymer matrix may thus be present. In the absence ofaggregates, there would be an increased degree of bonding betweenpolymers and biomass. Additionally, lack of aggregates would allow forthe movement and flexure of polymer molecules. This would explain theincrease in TS and SAB observed at the 50:50 PVA:chitosan ratio.

The DOS of leather samples, as illustrated in FIG. 17, was observed todecrease with an increase in the PVA concentration of samples. The ML ofsamples, as illustrated in FIG. 18, displayed an opposite trend. DOSvalues were three times as large in samples with chitosan as the solepolymer as compared to samples with PVA as the sole polymer. Withoutwishing to be bound by any particular theory, it is believed thatchitosan molecules have a higher affinity for water molecules thanmolecules of PVA, possibly due to the positive charge on the amine groupof chitosan at low pH. This charged amine group may also be more likelyto bind to other molecules, e.g. biomass particles, glycerol, or EPSconstituents, within the system. Due to the charged amine group,chitosan molecules may be more likely to remain bound during soaking,which would explain the lower ML values observed at higherconcentrations of Ch and vice versa.

EXAMPLE 9 Effect of Blending Time on Fungal Particle Length

40 grams of raw (unprocessed) fungal biomass and 40 mL of deionizedwater were placed into a small Oster blender and blended for 10 seconds.3 mL of the resulting mixture was removed from the blender and combinedwith 27 mL of deionized water to form 30 mL of a “10-second blend” testmaterial. The remaining mixture in the blender was blended for anadditional 10 seconds, and another 3 mL sample was removed and combinedwith 27 mL of deionized water to form 30 mL of a “20-second blend” testmaterial. This process was repeated with another 20 seconds of blendingto obtain a “40-second blend,” and again after still another 20 secondsof blending to obtain a “60-second blend.” Each of the test materialswas then further diluted 9:1 in deionized water to form four 300 mLsamples, each comprising 1 vol % of the blended mixture.

75 μL of each of the four 1 vol % samples was placed onto a microscopeslide, and photomicrographs of each sample were taken. In eachphotomicrograph, the apparent length of 30 fungal particles wasmeasured, and these apparent lengths were converted to the true lengthof each particle based on the magnification used in the microscope.Histograms of particle length for the blends are illustrated in FIGS.19A, 19B, 19C, and 19D, respectively.

EXAMPLE 10 Effect of Loading Ratio on Foaming During Manufacture

Raw (unprocessed) fungal biomass was chopped into pieces approximately 1cm square and added in varying amounts to each of several 400 mL beakerstogether with water, a PVA solution, chitosan, and adipic acid. Theheight of the mixture in each beaker was measured, and each mixture wasthen blended for one minute using a Hamilton Beach HB08 hand mixer,whereupon the height of the mixture was measured again. Each mixture wasthen stirred (large stir bar, 60 rpm) at 180° C. for 30 minutes, afterwhich the height of the mixture was measured a third time, and then afourth time after addition of a volume of acetic acid to the beaker; themixture was stirred for 10 additional minutes during cooling. Eachmixture was then poured into a flat tray, allowed to dry for two days atroom temperature, then removed from the tray and heat-pressed at 275° F.in silicone textured molds for ten minutes at a time, five separatetimes. The density of each heat-pressed sample was then measured. FIG.20 illustrates the “blend overrun,” “heating overrun,” and “overalloverrun”—respectively, the change in volume relative to the startingmixture after blending, heating, and acetic acid addition—as well as thedensity for each mixture as a function of the loading ratio.

EXAMPLE 11 Physical Properties of Fungal Leather Analogs—Size-Reducedvs. Intact Biomass

Eight samples of a fungal leather analog material were producedaccording to the method illustrated in FIG. 3 and the descriptionassociated therewith, except as otherwise noted. Of these eight samples,three were made from inactivated fungal biomass that was size-reducedprior to step 310—a first sample had no non-fungal textile backing, asecond sample had a non-fungal textile (cotton) backing on one side ofthe fungal layer, and a third sample had a non-fungal textile (cotton)layer “sandwiched” between two fungal layers. The other five sampleswere made from intact (non-size-reduced) biomats produced by a surfacefermentation process—fourth, fifth, and sixth samples had no non-fungaltextile backing, a seventh sample had a non-fungal textile (cotton)backing on one side of the fungal layer, and a third sample had anon-fungal textile (cotton) layer “sandwiched” between two fungallayers.

The size-reduced fungal biomasses were prepared as follows: water andthawed (previously frozen) processed biomass were added to a Vitamixblender in a 1:1 mass ratio. These were bended together forapproximately 2 minutes to produce a homogenous mixture of size-reducedbiomass in water. Separately, a solution of water, glycerol, chitosan,citric acid, and hydrochloric acid was prepared, with the respectivecomponents in a mass ratio of 200:17.5:6.3:1:13.5 respectively. Thetotal mass of solution was equal to that of the biomass-water blendedmixture. Once the chitosan was dissolved, the aqueous polymer solutionand the biomass-water mixture were combined. The newly formed mixturewas stirred under heat for approximately 30 min such that a homogenouspaste was formed. This paste was then cast into a flat nonstick tray andallowed to dry at ambient conditions. Once dried, the newly formed sheetmaterial was heat pressed at 100° C. for 10 minutes.

For those samples having non-fungal (cotton) layers, cotton backingmaterial was adhered to the sample using an aqueous solution of chitosan(1% w/v), citric acid (1% w/v), and hydrogen chloride (1% v/v). Thechitosan solution was painted onto appropriate side(s) of the fungallayer, and the cotton was applied to the wetted surface. The chitosanadhesive was allowed to dry for approximately 20 minutes, and the samplewas then heat-pressed at 275° F. for two minutes to bond the backingmaterial.

The eight fungal leather analog material samples were tested for ninephysical properties—thickness, tensile strength, tensile force,elongation at break, tear resistance, density, flexural rigidity, degreeof swelling, and mass loss after soaking. The results of these tests aregiven in Table 2 below.

TABLE 2 Intact biomass Size-reduced biomass No No No No TextileSandwiched textile textile textile Textile Sandwiched Parameter Unittextile backing textile #1 #2 #3 backing textile Thickness mm 1.4 2  2.5 0.4  0.85 1    0.61  0.90 Tensile MPa 6.5 6.3 9.8  3.65  1.75  3.16 4.93  3.31 strength Tensile N 54.6  75.6  147     8.83  8.86 18.9618.07 17.86 force Elongation % 17.8  20.5  18.3  17.45 13.79 20.53  9.0111.67 at break Tear N/mm 6.4 17.5  17.2   0.64 — — 48.57 31.90resistance Density g/cm³ 1.1 — —  1.39  1.26  1.51  1.11  1.42 Flexuralg · cm 10.59 — —  1.45  3.98  8.37  3.88  7.42 rigidity Degree of %49.33 — — 49.12 51.29 47.17 — — swelling Mass loss % 53.37 — — 49.6749.04 47.83 — — after soaking

EXAMPLE 12 Effect of Carbon-Nitrogen Ratio on Properties of FungalLeather Analog Materials

Four growth media for surface fermentation of fungal biomats (asdescribed in, e.g., the '050, 3 626, and '421 applications) wereprepared, each medium having an identical fructose content. The molarcarbon-to-nitrogen ratio (“CN ratio”) of each medium was adjusted byincreasing or decreasing the combined content of ammonium sulfate andurea (holding the ratio of these two components to each other constant)until the media had CN ratios of 5, 8.875, 10, and 20, respectively.Each medium was inoculated with 5% v/v of an MK7 inoculum via shakeflask inoculation.

250 mL of each inoculated medium was poured into each of four glasstrays, resulting in 16 total inoculated trays. The glass trays wereplaced in a wrapped reactor at a temperature of 27° C. and allowed toincubate for 120 hours, with photographs of each tray taken at 72, 96,and 120 hours. The biomass from each tray was then harvested andinactivated in deionized water at 70° C. for 30 minutes. The wet yieldof each sample was determined after inactivation to assess relativegrowth performance.

Each sample of biomass was then transformed into a fungal leather analogmaterial according to the method described in Example 11 above. Aftertanning, various physical parameters of each sample were measured. Theresults are given in Table 3 (the values shown are the average for eachCN ratio).

TABLE 3 CN ratio Parameter 5 8.875 10 20 Wet mass (g) 63.25 95.7 84.17582.5 Finished mass (g) 6.35 13.375 10.4 8.475 Thickness (mm) 0.565 1.1150.8975 0.6575 Density (g/cm³) 0.89101 1.07821 0.97116 0.94101 Tensilestrength 1.41537 1.71588 2.551 2.01343 (MPa) Elongation at 12.240912.5634 14.4092 12.9312 break (%) Tear resistance 1.13463 1.7848 1.214650.79766 (N/mm) Degree of 15.2407 22.5345 22.1204 16.9055 swelling (%)Mass loss (%) 55.6727 48.5781 50.6389 59.0071 Flexural rigidity 1.209365.03108 1.70845 0.44633 (g · cm)

Various qualitative distinctions between the samples were also observed,both before and after tanning process. Biomats grown on media having aCN ratio of 5 were more “slippery” and noticeably thinner in places,especially in portions of the biomat grown near the center of the tray;once inactivated, these fungal samples were extremely flexible. Biomatsgrown on media having CN ratios of 8.875 and 10 were very stiff afterinactivation, likely due to the thickness of the biomat. Biomats grownon media having a CN ratio of 20 were more flexible than those grown onmedia having CN ratios of 8.875 and 10, both before and after theinactivation step.

After the tanning process, it was observed that samples derived frommedia having a CN ratio of 5 had uneven thickness and were inflexible inthose areas where the material was thickest; areas that were undergreater pressure during the heat-pressing step were shinier and had asmoother texture, possibly due to the compression of the heat pressaligning and compacting filaments of the filamentous fungus. Samplesderived from media having a CN ratio of 8.875 were thickest and shrunkthe most during the drying step, had uneven surface textures, and feltstiffer than other samples. Samples derived from media having a CN ratioof 10 were intermediate in thickness and were more flexible than thesamples derived from media having a CN ratio of 8.875, but alsodisplayed an uneven surface; like the samples derived from media havinga CN ratio of 5, areas exposed to the greatest compression during theheat-pressing step were noticeably shinier. Samples derived from mediahaving a CN ratio of 20 were intermediate in thickness between the CNratio 5 and CN ratio 10 samples, and were relatively flexible and hadslightly more uniform surfaces; once again, those areas that were mostcompressed during heat-pressing were shiniest.

EXAMPLE 13 Thermal Doping of Fungal Leather Analogs

Each of five experimental samples was prepared as follows: 75 grams ofglycerol, 27 grams of chitosan, 4.3 grams of citric acid, 880milliliters of water, and 13.5 milliliters of concentrated hydrochloricacid were placed in a beaker and stirred until the chitosan dissolved.Separately, 80 grams of wet filamentous fungal biomass, produced by asurface fermentation method as described herein and in the '050, '626,and '421 applications, and 80 milliliters of water were placed in akitchen blender and blended until homogeneous. 7.2 grams of a thermaldopant was then added to the blender (except in the case of the controlsample), and the mixture was again blended until homogeneous. 160 gramsof the chitosan solution were then added to the blender, and thismixture was again homogenized; 300 grams of the resulting mixture waspoured into a small nonstick tray and dried at 90° F. for 23 hours. Thedried sample was heat-pressed at 100° C. for 10 minutes to produce aflat sheet of moderate flexibility approximately 2 millimeters thick.

Thermal properties of each of the samples were measured. The results ofthese measurements are given in Table 4; control samples of undoped hideleather, undoped blended fungal leather analog, and undoped leather madefrom intact biomats (CN ratios of 8.875, 10, and 20, denoted “CN8,”“CN10,” and “CN20,” respectively) were also tested for comparison.

TABLE 4 Thermal Thermal Volumetric Sample conductivity diffusivityspecific heat ID Dopant (W/m · K) (mm²/s) (MJ/m³ · K) LC None (hideleather 0.100 0.091 1.096 control) ALU Aluminum oxide 0.243 0.132 1.867CNT None (fungal 0.181 0.122 1.487 leather control) EVA Ethylene vinyl0.215 0.081 2.655 acetate Lig Lignin 0.199 0.139 1.443 YTT Yttrium oxide0.155 0.058 2.669 Bent Bentonite 0.200 0.054 3.704 CN8 None 0.234 0.1381.691 CN10 None 0.314 0.132 2.373 CN20 None 0.302 0.216 1.399

EXAMPLE 14 Effect of Polyvinyl Acetate on Material Performance

A large mixture of biomass, water, glycerol, and adipic acid was mixedin a blender and separated into five equal portions. Five separate 6%polymer solutions containing polyvinyl alcohol (PVA) and chitosan in an80:20 mass ratio were made, each solution containing a different type ofKuraray PVA. Each polymer solution was combined with a portion of thebiomass mixtures, thereby making five separate leather precursormixtures. Each of these leather precursor mixtures was individuallymixed using a handheld immersion blender and poured into a small Pyrextray, then dried at room temperature with a fan blowing over the trays.When each sample reached a moisture content of no more than 20%, it washeat-pressed at 100° C. for ten minutes, two separate times. The sampleswere then allowed to dry overnight at room temperature and subsequentlytested for tensile and tear strength and qualitatively examined fortexture and water resistance. Each leather sample had a loading ratio of75:25 and a plasticizer content of 22.5 wt %. Results of this testingare provided in Table 5.

TABLE 5 PVA Tensile Tear Type/ Molecule Degree of Viscosity StrengthAverage Strength Average Name Description Hydrolysis (mPa · s) Sample(N/mm²) (N/mm²) (N/mm) (N/mm) Texture Notes Water Resistance NotesExceval Copolymer 99.3 13.8  a 3.579 3.277 5.877 6.044 Smooth surface.Droplets bead up HR-3010 of PVA and b 2.837 6.210 Cracks on both on bothsides. PE. 80:20 c 3.414 sides when bent Minimal swelling. 180 degrees.Minimal discoloration. Low to Medium flexibility. Exceval Copolymer 98.127.8  a 3.574 3.505 5.788 7.215 Smooth surface. Droplets bead up RS-2117of PVA and b 3.548 7.870 Cracks on both on both sides. PE. 80:20 c 3.3947.987 sides when bent Minimal swelling. 180 degrees. Minimaldiscoloration. Lower flexibility. Exceval Copolymer 98.6 4.3 a 3.1453.356 5.315 5.447 Smooth surface. Droplets spread over AQ-4104 of PVAand b 3.073 6.016 Cracks on both surface on both sides. PE. 80:20 c3.849 5.009 sides when bent Relatively fast absorption. 180 degrees.Minimal discoloration. Lowest flexibility. Poval Pure PVA 98.8 27.3  a4.590 4.790 7.726 7.152 Smooth surface. Droplets bead up 28-98 S2 b4.330 7.558 Cracks on the on both sides. c 5.449 6.171 non-skin sideSlightly faster absorption. when bent Minimal swelling. 180 degrees.Minimal discoloration. Medium flexibility. Elvanol Pure PVA 99.6 29.1  a4.905 3.836 5.566 5.997 Smooth surface. Droplets bead up 71-30 b 3.5296.428 Cracks on both on both sides. c 3.073 sides when bent Minimalswelling. 180 degrees. Minimal discoloration. Low to Medium flexibility.

The viscosity of the PVA, which correlates directly with molecularweight, had a significant effect on tensile and tear strength, withlow-viscosity PVAs resulting in poor tensile and tear properties. Forhigher-viscosity PVA types, the degree of hydrolysis appeared to be thedetermining factor for tensile and tear properties; samples with a lowdegree of hydrolysis displayed better tensile and tear properties.Without wishing to be bound by any particular theory, the presentinventors hypothesize that a higher concentration of acetate groups actsas a plasticizer, allowing for free movement of internal molecules andreduced cracking and brittleness on a microscopic level, therebyincreasing the overall strength and flexibility of samples.

The present disclosure, in various aspects, embodiments, andconfigurations, includes components, methods, processes, systems and/orapparatus substantially as depicted and described herein, includingvarious aspects, embodiments, configurations, sub-combinations, andsubsets thereof. Those of skill in the art will understand how to makeand use the various aspects, aspects, embodiments, and configurations,after understanding the present disclosure. The present disclosure, invarious aspects, embodiments, and configurations, includes providingdevices and processes in the absence of items not depicted and/ordescribed herein or in various aspects, embodiments, and configurationshereof, including in the absence of such items as may have been used inprevious devices or processes, e.g., for improving performance,achieving ease and\or reducing cost of implementation.

The foregoing discussion of the disclosure has been presented forpurposes of illustration and description. The foregoing is not intendedto limit the disclosure to the form or forms disclosed herein. In theforegoing Detailed Description for example, various features of thedisclosure are grouped together in one or more, aspects, embodiments,and configurations for the purpose of streamlining the disclosure. Thefeatures of the aspects, embodiments, and configurations of thedisclosure may be combined in alternate aspects, embodiments, andconfigurations other than those discussed above. This method ofdisclosure is not to be interpreted as reflecting an intention that theclaimed disclosure requires more features than are expressly recited ineach claim. Rather, as the following claims reflect, inventive aspectslie in less than all features of a single foregoing disclosed aspects,embodiments, and configurations. Thus, the following claims are herebyincorporated into this Detailed Description, with each claim standing onits own as a separate preferred embodiment of the disclosure.

Moreover, though the description of the disclosure has includeddescription of one or more aspects, embodiments, or configurations andcertain variations and modifications, other variations, combinations,and modifications are within the scope of the disclosure, e.g., as maybe within the skill and knowledge of those in the art, afterunderstanding the present disclosure. It is intended to obtain rightswhich include alternative aspects, embodiments, and configurations tothe extent permitted, including alternate, interchangeable and/orequivalent structures, functions, ranges or steps to those claimed,whether or not such alternate, interchangeable and/or equivalentstructures, functions, ranges or steps are disclosed herein, and withoutintending to publicly dedicate any patentable subject matter.

The invention claimed is:
 1. A method for preparing a durable sheetmaterial comprising fungal biomass, comprising: (a) causing an aqueouspolymer solution, comprising a solvent and a polymer, to infiltrate aninactivated fungal biomass to a fungal biomass:polymer loading ratio ofbetween 25:75 and 75:25; and (b) curing the biomass to remove solventfrom the biomass and form the durable sheet material, wherein theinactivated fungal biomass comprises a cohesive fungal biomass.
 2. Themethod of claim 1, further comprising at least one of (i) adding athermal dopant to the inactivated fungal biomass and (ii) adding athermal dopant to the durable sheet material after step (b).
 3. Themethod of claim 2, wherein the thermal dopant is selected from the groupconsisting of a ceramic material, a metallic material, a polymericmaterial, and combinations thereof.
 4. The method of claim 2, whereinthe thermal dopant is selected from the group consisting of activatedcharcoal, aluminum oxide, bentonite, diatomaceous earth, ethylene vinylacetate, lignin, nanosilica, polycaprolactone, polylactic acid,silicone, and yttrium oxide.
 5. The method of claim 1, wherein thefungal biomass comprises fungal mycelia.
 6. The method of claim 1,wherein step (a) comprises agitating the inactivated fungal biomass andthe solution together for a time period.
 7. The method of claim 6,wherein the time period is selected from the group consisting of atleast about 4 hours, at least about 5 hours, at least about 10 hours, atleast about 15 hours, at least about 20 hours, and at least about 25hours.
 8. The method of claim 6, wherein the time period is betweenabout 10 hours and about 20 hours.
 9. The method of claim 6, wherein theagitating is carried out at a pressure other than atmospheric pressure.10. The method of claim 1, wherein the cohesive fungal biomass isproduced by a surface fermentation process or a submerged solid surfacefermentation process.
 11. The method of claim 1, further comprisingsubjecting the inactivated fungal biomass to treatment with at least onechemical selected from the group consisting of calcium hydroxide andtannins.
 12. The method of claim 1, wherein the polymer is selected fromthe group consisting of polyvinyl alcohol, chitosan, polyethyleneglycol, alginates, starches, polycaprolactones, polyacrylic acids,hyaluronic acid, and combinations thereof.
 13. The method of claim 1,wherein the solution further comprises a crosslinker selected from thegroup consisting of citric acid, tannic acid, suberic acid, adipic acid,succinic acid, extracted vegetable tannins, glyoxal, and combinationsthereof.
 14. The method of claim 1, wherein the solution furthercomprises a plasticizer selected from the group consisting of glyceroland esters thereof, polyethylene glycol, citric acid, oleic acid, oleicacid polyols and esters thereof, epoxidized triglyceride vegetable oils,castor oil, pentaerythritol, fatty acid esters, carboxylic ester-basedplasticizers, trimellitates, adipates, sebacates, maleates, biologicalplasticizers, and combinations thereof.
 15. The method of claim 1,wherein the inactivated fungal biomass comprises fungal mycelia producedby submerged fermentation.
 16. The method of claim 15, wherein thefungal mycelia produced by submerged fermentation are in the form of apaste.
 17. The method of claim 1, wherein the aqueous polymer solutionfurther comprises at least one of a pigment, a solubilizer, and a pHadjusting agent.
 18. The method of claim 17, wherein the solutioncomprises a solubilizer selected from the group consisting ofhydrochloric acid, acetic acid, formic acid, lactic acid, andcombinations and mixtures thereof.
 19. The method of claim 17, whereinthe solution comprises a pH adjusting agent selected from the groupconsisting of hydrochloric acid, acetic acid, formic acid, lactic acid,and combinations and mixtures thereof.
 20. The method of claim 1,wherein the durable sheet material comprises proteins crosslinked withisopeptide bonds.
 21. The method of claim 1, wherein the inactivatedfungal biomass comprises a biomat, or portion thereof, produced by asurface fermentation process.
 22. The method of claim 1, wherein theinactivated fungal biomass was grown on a growth medium with acarbon-to-nitrogen molar ratio between about 5 and about 20, or betweenabout 7 and about 15.